Genome Editing

Germline genetic intervention (GGI) has been one of the most discussed topics within the bioethics literature since 2012, when the programming of CRISPR/Cas9 for a specifically targeted gene region has become possible. While some authors are optimistic about what GGI may offer, others strongly disagree and refute the use of this technology for different reasons. This paper will aim to examine one of the most widespread arguments against GGI, namely "heritability" argument, comprehensively. Firstly, it will aim to examine the moral importance of the germline. Secondly, it will try to understand three possible assumptions of the heritability argument. Then it will try to respond to these assumptions and argue that they are neither scientifically supportable nor rationally solid for rejecting GGI altogether.

Transgenic science and technology are fundamental to
the state-of-the-art plant molecular genetics and crop
improvement. The new generation of technology endeavors
to introduce genes ‘stably’ into ‘site-specific’ locations and
in ‘single copy’ without the integration of extraneous vector
‘backbone’ sequences or ‘selectable markers’. Numerous plant
transformation technologies have developed with the aim
of achieving these objectives. Here we discuss some of these
technologies, which can push the development of ‘better
transgenic plants with desirable characters only’.

Development of new plant breeding techniques have facilitated easy manipulation of plants at genetic level. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated protein9 (Cas9) system is a valuable addition in programmable nucleases. The CRISPR/Cas9 system uses an RNA component to recognize a target DNA sequences and it has shown promising results with respect to simultaneous editing of multigenic plant traits. In this review, components of CRISPR/Cas9, their construction and its methods of delivery to plant cells are analyzed. Variation in nucleotide sequence of the protospacer adjacent motif, codon optimization and progress in web-based bioinformatic tools, will make CRISPR/Cas9 systems more efficient for plants. Development and optimization of protocols to efficiently targ et  all plant species is still under development. Along with this, methods to inspect induced mutation and efficiency of the system have also been reviewed. Auxiliary improvement...

The CRISPR/Cas9 system is now commonly employed for genome editing in various plants such as Arabidopsis, rice and tobacco. In general, in genome editing of the Arabidopsis genome, the SpCas9 and guide RNA genes are introduced into the genome by the floral dip method. Mutations induced in the target sequence by SpCas9 are confirmed after selecting transformants by screening the T 1 seed population. The advantage of this method is that genome-edited plants can be isolated easily. However, mutation efficiency in Arabidopsis using SpCas9 is not as high as that achieved in rice and tobacco, which are subjected to a tissue culture step. In this study, we compared four promoters and found that the parsley UBIQITIN promoter is highly active in Arabidopsis meristem tissue. Furthermore, we examined whether a simple heat treatment could improve mutation efficiency in Arabidopsis. Just one heat treatment at 37 • C for 24 h increased the mutation efficiency at all four target sites from 3 to 42%, 43 to 62%, 54 to 75% and 89 to 91%, without detectable off-target mutations. We recommend heat treatment of plate-grown plants at 37 • C for 24 h as a simple method to increase the efficiency of CRISPR/Cas9-mediated mutagenesis in Arabidopsis.

In den letzten fünf Jahren ist ein wachsendes Interesse an den neuen Möglichkeiten der Genom-Editierung zu erkennen. So zeigen sich nicht nur eine rasant steigende Zahl an Studien und wissenschaftlichen Publikation zu CRISPR und Co., die Anwendungsmöglichkeiten der Genom-Editierung werden auch breit in den öffentlichen Medien verhandelt. Zahlreiche Magazine, darunter Der Spiegel, The Economist, National Geographic oder auch TIME haben dem Thema mittlerweile eine eigene Titelseite gewidmet. Die Rezeption in Wissenschaft und den Medien ist dabei von moralische Ambiguität gekennzeichnet. Während regelmäßig darauf hingewiesen wird, dass die neuen und zukünftigen Möglichkeiten der Genom-Editierung eine ,schnelle, präzise, einfache und kostengünstige' Möglichkeit zur Veränderung von Lebewesen darstellen, wird zugleich eine Warnung hinterhergeschoben: zwar biete die Genom-Editierung in der klinischen Anwendung große Potenziale bezüglich somatischer Gentherapien. Die neuen Werkzeuge dürften allerdings nicht zur Veränderung der Keimbahn dienen, zumindest nicht, um mit entsprechend veränderten Embryonen oder Keimbahnzellen eine Schwangerschaft herbeizuführen. Exemplarisch sind in diesem Zusammenhang die zwei weit rezipierten Kommentare in Nature und Science zu nennen, die kurz vor der Veröffentlichung der ersten Versuche an menschlichen Embryonen durch ein chinesisches Forscherteam veröffentlich wurden. 1 Die Sorgen um die möglichen Gefahren der Genom-Editierung kulminieren dabei in der Behauptung, dass entsprechende Techniken zur Veränderung der Keimbahn zu einer Form der Eugenik führten. So findet sich in der Reihe an Magazinen, die den Durchbrüchen der Genom-Editierung eine Titelgeschichte widmeten, auch The Spectator wieder. Der dort eingeschlagene Ton ist düster: " Eugenics is back " lesen wir, bebildert von Föten und einem Säugling in einer Petrischale. 2 Diese Sorge einer Rückkehr oder auch Erneuerung ,der Eugenik' zieht sich nicht nur durch die journalistische Presse. Die Wissenschaftshistoriker und Eugenik-Forscher Daniel J. Kevles und Nathaniel C. Comfort haben sich im Zuge der Auseinandersetzung mit CRISPR um eine Parallelisierung mit der vergangenen Eugenik bemüht 3 und auch in Science wurde ein entsprechender Vergleich aufgegriffen. Kurz bevor das Magazin CRISPR zur technischen Entwicklung des Jahres 2015 prämierte, druckte es einen besorgten

Genome editing is an important tool for gene functional studies as well as crop improvement. The recent development of the CRISPR/Cas9 system using single guide RNA molecules (sgRNAs) to direct precise double strand breaks in the genome has the potential to revolutionize agriculture. Unfortunately, not all sgRNAs are equally efficient and it is difficult to predict their efficiency by bioinformatics. In crops such as cotton (Gossypium hirsutum L.), with labor-intensive and lengthy transformation procedures, it is essential to minimize the risk of using an ineffective sgRNA that could result in the production of transgenic plants without the desired CRISPR-induced mutations. In this study, we have developed a fast and efficient method to validate the functionality of sgRNAs in cotton using a transient expression system. We have used this method to validate target sites for three different genes GhPDS, GhCLA1, and GhEF1 and analyzed the nature of the CRISPR/Cas9-induced mutations. In our experiments, the most frequent type of mutations observed in cotton cotyledons were deletions (∼64%). We prove that the CRISPR/Cas9 system can effectively produce mutations in homeologous cotton genes, an important requisite in this allotetraploid crop. We also show that multiple gene targeting can be achieved in cotton with the simultaneous expression of several sgRNAs and have generated mutations in GhPDS and GhEF1 at two target sites. Additionally, we have used the CRISPR/Cas9 system to produce targeted gene fragment deletions in the GhPDS locus. Finally, we obtained transgenic cotton plants containing CRISPR/Cas9-induced gene editing mutations in the GhCLA1 gene. The mutation efficiency was very high, with 80.6% of the transgenic lines containing mutations in the GhCLA1 target site resulting in an intense albino phenotype due to interference with chloroplast biogenesis.

Genome editing is a method to change the DNA inside an organism. It allows the scientists to remove, to alter, or to add the DNA sequence in particular location in genome. Nowadays, one of the most known genome editing technologies is CRISPR-Cas9. CRISPR-Cas9 stands for Clustered Regularly Interspaced Palindromic Repeats and the protein Cas9 (CRISPR-associated). It is a RNA guided and cleaves the DNA double strand, providing a powerful method to induce deletions, insertions and specific sequence changes at defined target sites. It has generated many research and inventions because it makes possible to correct errors in the genome, and turn on/off genes in cells and organisms quickly, cheaply, accurately and efficiently than other exsisting genome editing methods.

Crisper/Cas9 is one of the latest and most efficient genome editing tools which is in fact is the part of bacteria and archaea adaptive immunity. Crisper/Cas9 comprises of the guide RNA for targeted binding and Cas9 (endonuclease) for cleavage which attack sequences adjacent to PAM. Adeno associated viruses are mostly used to of deliver CRISPER/Cas9. Several researches targeted the undesirable sequences to manipulate gene expression in different organisms (zebrafish, mouse, etc.) and induced pluripotent cells (iPSCs). This simple mechanism has been applied in understanding and treatment of several complex diseases. Recent advances have expanded its potential which are discussed in detail in this paper. The biggest challenge is off-targeting. It has many ethical considerations as well. Despite the limitations, CRISPER/Cas9 has revolutionised the medical field with its wide applications. This paper 1st discusses how does the crisper/cas9 works in bacteria and as a gene manipulation tool. It then discusses how it is designed and delivered most effectively; followed by its applications and challenges.

Plants and microbes are co-evolved and interact with each other in nature. Plant-associated microbes, often referred to as plant microbiota, are an integral part of plant life. Depending on the health effects on hosts, plant-microbe (PM) interactions are either beneficial or harmful. The role of microbiota in plant growth promotion (PGP) and protection against various stresses is well known. Recently, our knowledge of community composition of plant microbiome and significant driving factors have significantly improved. So, the use of plant microbiome is a reliable approach for a next green revolution and to meet the global food demand in sustainable and eco-friendly agriculture. An application of the multifaceted PM interactions needs the use of novel tools to know critical genetic and molecular aspects. Recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-mediated genome editing (GE) tools are of great interest to explore PM interactions. A systematic understanding of the PM interactions will enable the application of GE tools to enhance the capacity of microbes or plants for agronomic trait improvement. This review focuses on applying GE techniques in plants or associated microbiota for discovering the fundamentals of the PM interactions, disease resistance, PGP activity, and future implications in agriculture.

While CRISPR-Cas genome editing technology has been heralded as a great ad- vancement for science, human health, bioengineering and medicine, there is a potential dark side of CRISPR genome editing that may be a greater danger to humanity than nuclear weapons. We present some of the more obvious poten- tial military and bioterrorism applications of CRISPR and related genome editing technology. CRISPR weapons have significant advantages over conventional nu- clear weapons. The balance of power based on mutual assured destruction cannot be maintained once CRISPR weapons enter the international arena. Disturbingly, because of the easy entrance to the technology, it opens the door for small organi- zations and countries to enter the international arms arena. The simplicity and ease of development of such weapons makes it essential that CRISPR genome editing technology be carefully applied and regulated.

This paper explores the ethics of introducing genome-editing technologies as a new reproductive option. In particular, it focuses on whether genome editing can be considered a morally valuable alternative to preimplantation genetic diagnosis (PGD). Two arguments against the use of genome editing in reproduction are analysed, namely safety concerns and germline modification. These arguments are then contrasted with arguments in favour of genome editing, in particular with the argument of the child’s welfare and the argument of parental reproductive autonomy. In addition to these two arguments, genome editing could be considered as a worthy alternative to PGD as it may not be subjected to some of the moral critiques moved against this technology. Even if these arguments offer sound reasons in favour of introducing genome editing as a new reproductive option, I conclude that these benefits should be balanced against other considerations. More
specifically, I maintain that concerns regarding the equality of access to assisted reproduction and the allocation of scarce resources should be addressed prior to the adoption of genome editing as a new reproductive option.

The interest in exploring the interplay of genomics and Islamic ethics took an important turn at the beginning of the 1990s, when the international Human Genome Project was declared. Since then, both Muslim religious scholars and biomedical scientists have been examining the relevant ethical questions from an Islamic perspective in addition to providing recommendations for policy-making pertinent to biomedical and genetic research in the Muslim world.

The Portuguese Society of Human Genetics (SPGH - Sociedade Portuguesa de Genética Humana) was founded in 1996 by a group of 68 medical and clinical laboratory geneticists. Its mission, statutes and governance resemble those of societies of similar ilk. Aligned with these and with the European Society of Human Genetics, so too the SPGH has organized annual meetings, constituting a major event that brings together professionals and students from all over the Country. In 2018, from the 15th to the 17th November, Porto hosted the SPGH's 22nd Annual Meeting. Recent developments in several fields of human genetics were brought to discussion, with the spotlight on epigenetic regulation and dysregulation, the revolutionary role of immunotherapy in cancer treatment, advances in etiologic diagnosis and therapy of neurological disorders, as well as the ethical implications of genome editing. These were addressed in the format of plenary lectures, thematic sessions or Panel discussions (detailed proceedings available at www.spgh/net/reuniao-anual/22a-reuniao/). The programme included several Satellite meetings and three sessions each of oral and poster presentations, selected from a total of 97 Abstracts. The high scientific standard was set by the excellence of the guest speakers, all renowned experts in the respective fields, while the vivid discussions, exchange of ideas and new collaborations are a measure of the meeting's success. The event was rounded off with the usual SPGH prize-giving ceremony, with five awards: best presentations in Basic Research, Clinical Research, Clinical Case Reports, the Amândio Tavares Young Scientist Award and the ESHG Fellowship Award.

CRISPR/Cas 9 is a newly developed and naturally occurred genome editing tool, which is originally used by bacteria for immune defence. In the past years, it has been quickly employed and modified to precisely edit genome sequences in both plants and animals. Compared with the well-developed zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), CRISPR/Cas 9 has lots of advantages, including easier to design and implement, higher targeting efficiency, and less expensive. Thus, it is becoming one of the most powerful tools for knockout of an individual gene as well as insertion of one gene and/or control of gene transcription. Studies have shown that CRISPR/Cas 9 is a great tool to edit many genes in a variety of plant species, including the model plant species as well as agriculturally important crops, such as cotton, maize and rice. CRISPR/Cas 9-based genome editing can be used for plant functional studies and plant improvement to yield, quality and tolerance to environmental stress

While CRISPR/Cas9 has become a lightning rod for fears about humanity’s increasing capacity to engineer biological life, the mainstream of Anglo-American bioethics struggles to discern much wrong with genome editing of human beings in vitro. In this paper, we analyze the notion of biopolitics and consider what contribution it may make to debates on genome editing. We disambiguate the different senses of these two key terms: ‘biopolitics’, and ‘life’, and try to show how particular authors in the biopolitics literature draw on and emphasize different versions of these concepts. In the final section of the paper, we venture some suggestions as to the contribution that a number of these approaches might make to moving beyond a focus on risk and individual liberty in order to address the urgent bioethical questions surrounding the use of CRISPR/Cas9 to edit the human genome.

Fragrant rice is favoured worldwide because of its agreeable scent. The presence of a defective badh2 allele encoding betaine aldehyde dehydrogenase (BADH2) results in the synthesis of 2-acetyl-1-pyrroline (2AP), which is a major fragrance compound. Here, transcription activator-like effector nucleases (TALENs) were engineered to target and disrupt the OsBADH2 gene. Six heterozygous mutants (30%) were recovered from 20 transgenic hygromycin-resistant lines. Sanger sequencing confirmed that these lines had various indel mutations at the TALEN target site. All six transmitted the BADH2 mutations to the T1 generation; and four T1 mutant lines tested also efficiently transmitted the mutations to the T2 generation. Mutant plants carrying only the desired DNA sequence change but not the TALEN transgene were obtained by segregation in the T1 and T2 generations. The 2AP content of rice grains of the T1 lines with homozygous mutations increased from 0 to 0.35-0.75 mg/kg, which was similar to...

Genome editing, or genome editing with
engineered nucleases, is a technology that, using
engineered nucleases, allows site-specific single-base
mutations or the insertion, deletion or replacement of
DNA sequences in a specific site in the genome of an
organism. Genome editing is based on the induction
of double strand breaks (DSBs) in the DNA in the
locus of interest to introduce mutations in that locus.
In fact, after DSB induction, the damage will be
repaired by processes (the non-homologous end joining
and/or the homology-directed repair), that occur
naturally in the cells and during which mutations may
occur. DSBs can be induced by different nucleases,
all capable of specifically recognising a locus in the
genome. The most promising is the CRISPR/Cas system,
for ease of designing nucleases with sequence
specificity and for the fact that it can be used in nearly
every organism. In the CRISPR/Cas9 system, the
recognition of the DNA sequence to be modified is
operated by an RNA sequence. After successful DNA
DSB, the cell proceeds with the repair of DNA.
Generally, the cell uses non-homologous end joining,
which produces substitutions, insertions and deletions
of nucleotides in the damaged DNA site, and usually
leads to loss of function of the target gene. When
using this mode, the genome editing can be considered
a biological site-specific mutagenesis, different
from the mutagenesis induced by physical or chemical
agents which randomly induce mutations through the
entire genome. On the contrary, when homologydirected
repair is involved, genome editing can be
considered a predetermined biological mutagenesis
that modifies or corrects the target gene in the sense
determined by the investigator. Applying genome editing
to plants requires also ancillary technologies,
according to the species and cell types. First, in vitro
culture techniques, especially protoplast cultures,
might be necessary for the production of cells that can
be subjected to the nuclease treatment. Then, transformation
vectors (Agrobacterium, viruses or biolistic
methods) are needed to enable the transfer of the
components required for genome editing to the plant
cell. The vectors may be stable or transient; in the latter
case, both the possible cytotoxicity of constitutively
expressed nucleases and the production of transgenic
plants would be avoided. Concerning the first results
obtained using this technology, mutations in target
genes of cultivated plants were obtained mostly
through non-homologous end joining for traits related
to morphology, quality and to the resistance to
pathogens and herbicides, in both herbaceous and
woody species. Results were also reported exploiting
the homology-directed repair. Overall, the genome
editing technology proved suitable to introduce precise
and predictable gene mutations directly into elite
cultivars, reducing the duration of traditional crossing
and backcrossing breeding, with the possibility to
modi fy more than one genes per experiment .
Although many advances in genome editing technology
have been achieved in recent years, some technical
problems remain to be solved, including the need
for increasing the efficiency of the system, the production
of off-target mutations, the influence of chromatin
structure on the editing efficiency, the possible side
effects on genes lying close to target genes and the
efficiency of the technology in polyploid species
(where many copies of target genes occur). In conclusion,
the CRISPR/Cas system has emerged as the
most important tool for the future of genetics because
of its simplicity, versatility and efficiency. It will have a
major impact on both basic and applied research and
will be used to produce cultivars with improved disease
resistance, with a higher nutritional value, and
able to survive climate changes, more suitable as
bioenergy crops, producing useful chemicals and biomolecules.

This paper sets out to provide an overview of the ethical issues discussed in the debate on gene editing technology CRISPR Cas9. It shows how the debate has been polarised between positive and negative reactions from the scientific and bioethical communities how these reactions are not new in bioethics, but they have become “patterned and institutionalised”. The paper looks critically at the focus on human embryos and on germline modification and shows how other applications of the technology, albeit largely overlooked by the debate, are ethically relevant and deserve attention.

We, Other Utopians… Recombinant DNA, Genome Editing, and Artificial Life is analyzing the topics of genome editing/ recombinant DNA on the basis of ethnographic research in the post-communist context. The book focuses on the topics of human DNA editing and genome repair on two levels. First, inspired by texts exploring the concept of life and the body in general, it conceptually and analytically works with various approaches to engineered life and embodiments from the perspective of anthropology, sociology, and science and technology studies. Second, it presents an analysis of artificial life, and biotechnological embodiments on concrete technologies – genome editing, and recombinant DNA. The book explores the theme of genome editing based on ethnographic research conducted at a biochemical laboratory in the Czech Republic. The fieldwork was carried out from 2017 to 2019, mainly in a lab focusing on DNA damages and genomic risk of complex diseases or genetic vulnerabilities like breast cancer, infertility, ageing. Recombinant DNA is understood here as the exchange of DNA strands to produce and design new nucleotide sequence arrangements to heal or enhance human bodies and health in the future. Healing, prolonging, and enhancing future life are very often the aim of these manipulations and experiments. The book will examine the following issues: 1) analysis of body and biotechnological embodiments/materialities in the laboratory (conceptual work); and 2) biopolitical strategies using visions of future life and various ideas of utopia. The main analytical line of the book is based on original work on DNA repair and covers fieldwork analysis, including huge data sets, field notes, documents, text analysis, ethnographic talks, and in-depth interviews.
https://www.routledge.com/We-Other-Utopians-Recombinant-DNA-Genome-Editing-and-Artificial-Life/Slesingerova/p/book/9780367608019

Ein Supermarkt im Museum? Was bizarr klingt, wurde in Berlin Wirklich- keit: Für zehn Wochen (7. März bis 14. Mai 2019) hatten Besuchende
des Museums für Naturkunde Berlin die Möglichkeit, einen poppigen, einen artifiziellen Supermarkt im neu geschaffenen Experimentierfeld zu besuchen. Dieses Experimentierfeld stellt innerhalb des Museums den Ort dar, an dem sich Theorie und Praxis besonders eindrücklich miteinander verquicken. Der Name des Supermarkts: ErbUndGut. In ihm ließ sich zwar kein Abendessen kaufen, er bot aber stattdessen etwas anderes: Einen Blick hinter die Kulissen des spannenden, komplexen und streitbaren Themas Pflanzenzüchtung.

The complex allotetraploid genome is one of major challenges in cotton for repressing gene expression. Developing site-specific DNA mutation is the long-term dream for cotton breeding scientists. The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/ Cas9) system is emerging as a robust biotechnology for targeted-DNA mutation. In this study, two sgRNAs, GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2, were designed in the identical genomic regions of GhMYB25-like A and GhMYB25-like D, which were encoded by cotton A subgenome and the D subgenome, respectively, was assembled to direct Cas9-mediated allotetraploid cotton genome editing. High proportion (14.2–21.4%) CRISPR/Cas9-induced specific truncation events, either from GhMYB25-like A DNA site or from GhMYB25-like D DNA site, were detected in 50% examined transgenic cotton through PCR amplification assay and sequencing analyses. Sequencing results also demonstrated that 100% and 98.8% mutation frequency were occurred on GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2 target site respectively. The off-target effect was evaluated by sequencing two putative off-target sites, which have 3 and 1 mismatched nucleotides with GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2, respectively; all the examined samples were not detected any off-target-caused mutation events. Thus, these results demonstrated that CRISPR/Cas9 is qualified for generating DNA level mutations on allotetraploid cotton genome with high-efficiency and high-specificity. Cotton fiber and its derivative products play crucial roles in our daily life and the world economy, which had been estimated to directly determine the annual income of almost 100 million families from approximately 150 countries 1. Its annually worldwide economic impact had been assessed at approximately US$500 billion 1,2. The widely cultivated cotton cultivars are allotetraploid species, which consist of two set of subgenomes, " A subgenome " and " D subgenome ". Those two subgenomes cotton species reunited geographically by the transoceanic dispersal happened approximately 1–2 million years ago (MYA) 3. This polyploidization confers many excellent properties on tetraploid cotton, including longer cotton fiber length and higher cotton fiber strength, which make it possible to cultivate modern spinnable cotton cultivars. However, the complex genome feature of allotetraploid cotton presents a new challenge for cotton genes functional analyses and the genetic improvement through transgenic approach. Previously, many cotton genes were identified to be implicated in cotton fibre development 4–16 , stress responsees 17–20 , and pathogen immune regulation through expressed sequence tag (EST)-based cDNA library coupled with conventional RNAi and gene overexpression strategies 21–24. With the rapid development of high throughput deep sequencing technologies and bioinformatics, many diploid and allotetraploid cotton species were sequenced and assembled within recent years, including D-genome diploids cotton Gossypium raimondii (DD; 2n = 26) 25,26 , A-genome diploids Gossypium arboreum (AA; 2n = 26) 27 , ant the AtDt allotetraploid cotton species Gossypium hirsutum and Gossypium barbadense 28–30. Those excellent contributions extremely facilitate gene identification of interest and subsequent vector construction for functional genes analyses and screening via 'genotype-to-phenotype' approach. Thus, the arsenal of cotton genomic manipulation urgently require to be updated to meet the demand for rapid and precise dissecting gene functional analyses. Extensive studies have shown that high-frequent creating DNA double-strand breaks (DSBs) in desired nuclear DNA sites is a reliable approache to induce gene mutations 31. DSBs can trigger two distinct endoge-nous DNA repair mechanisms, error-prone nonhomologous end joining (NHEJ) and homology-directed repair

The central dogma (CD) of molecular biology constitutes the transfer of genetic information from DNA to RNA to protein. Major CD processes governing genetic flow include the cell cycle, DNA replication, chromosome packaging, epigenetic changes, transcription, posttranscriptional alterations, translation, and posttranslational modifications. The CD processes are tightly regulated in plants to maintain their genetic integrity throughout the life cycle as well as to pass the genetic material to the next generation. Engineering of various CD processes involved in gene regulation will accelerate crop improvement to feed the growing world population. CRISPR technology enables programmable editing of CD processes to alter DNA, RNA, or protein, which would have been impossible in the past. Here, an overview of recent advancements in CRISPR tool development and CRISPR-based CD modulations that expedite basic and applied plant research is provided. Furthermore, CRISPR applications in major thriving areas, such as gene discovery (allele mining and cryptic gene activation), introgression (de novo domestication and haploid induction), and application of desired traits beneficial to farmers or consumers (biotic/abiotic stress-resilient crops, plant cell factories, and delayed senescence), are described. Finally, the global regulatory policies, challenges, and prospects for CRISPR-mediated crop improvement are summarized.

In July 2018, the Nuffield Council of Bioethics released its long-awaited report on heritable genome editing (HGE). The Nuffield report was notable for finding that HGE could be morally permissible, even in cases of human enhancement. In this paper, we summarise the findings of the Nuffield Council report, critically examine the guiding principles they endorse and suggest ways in which the guiding principles could be strengthened. While we support the approach taken by the Nuffield Council, we argue that detailed consideration of the moral implications of genome editing yields much stronger conclusions than they draw. Rather than being merely ’morally permissible’, many instances of genome editing will be moral imperatives.

In questo contributo si intende discutere alcuni degli aspetti etici che la tecnica di germline genome editing con CRISPR/Cas solleva. Dopo aver presentato tale tecnica e aver circoscritto l’analisi ai trattamenti per risolvere malattie genetiche, si sosterrà che essa potrebbe condurre l’essere umano verso un terzo decentramento che renderebbe necessaria un’analisi interdisciplinare approfondita di tale pratica. In seguito, verranno analizzati alcuni argomenti utilizzati nel dibattito accademico per screditare la pratica di germline genome editing. Si tratteranno pertanto gli argomenti della “naturalezza della pratica” e del “consenso” sostenendo che essi non riescano a fornire ragioni definitive per considerare eticamente inaccettabile l’utilizzo di CRISPR al fine di modificare embrioni precoci e gameti sessuali. Successivamente si indagherà l’argomento della “sicurezza” sostenendo che CRISPR non possa dirsi una pratica sufficientemente sicura per essere  attualmente utilizzata. Inoltre, dopo aver analizzato il principio di precauzione, si riterrà opportuna una revisione delle misure e dei principi che regolano la sperimentazione riguardo a tale tecnica. Sulla base di queste considerazioni, si sosterrà che la tecnica di genome editing con CRISPR sulla linea germinale non sia da considerarsi una tecnica attualmente applicabile in un contesto terapeutico, sia da un punto di vista scientifico sia da un punto di vista etico. Tuttavia, qualora le questioni aperte dovessero in futuro essere risolte ci saranno valide ragioni per rendere disponibile la pratica per applicazioni terapeutiche.

Tendencies and efforts have shifted from genome description, DNA mapping, and DNA sequencing to active and profound reprogramming , repairing life on genetic and molecular levels in some parts of contemporary life science research. Mirroring and materializing this atmosphere, various life engineering technologies have been used and established in many areas of life sciences in the last decades. A contemporary progressive example of one such technology is DNA editing. Novel developments related to reproductive technologies, particularly embryo editing, prenatal human life engineering, and germline engineering need to be analyzed against the broader social and structural background. The crucial analytical scope for this paper is a specific field: the

Human pluripotent stem cells comprise induced pluripotent and embryonic stem cells, which have tremendous potential for biological and therapeutic applications. The development of efficient technologies for the targeted genome alteration of stem cells in disease models is a prerequisite for utilizing stem cells to their full potential. Genome editing of stem cells is possible with the help of synthetic nucleases that facilitate site-specific modification of a gene of interest. Recent advances in genome editing techniques have improved the efficiency and speed of the development of stem cells for human disease models. Zinc finger nucleases, transcription acti-vator-like effector nucleases, and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system are powerful tools for editing DNA at specific loci. Here, we discuss recent technological advances in genome editing with site-specific nucleases in human stem cells. Keywords Induced pluripotent stem cells Á Embryonic stem cells Á Zinc finger nucleases Á Transcription activator-like effector nucleases Á Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system

CRISPR system is a powerful defense mechanism in bacteria and archaea to provide immunity against viruses. Recently, this process found a new application in intended targeting of the genomes. CRISPR-mediated genome editing is performed by two main components namely single guide RNA and Cas9 protein. Despite the enormous data generated in this area, there is a dearth of high throughput resource. Therefore, we have developed CrisprGE, a central hub of CRISPR/Cas-based genome editing. Presently, this database holds a total of 4680 entries of 223 unique genes from 32 model and other organisms. It encompasses information about the organism, gene, target gene sequences, genetic modification, modifications length, genome editing efficiency, cell line, assay, etc. This depository is developed using the open source LAMP (Linux Apache MYSQL PHP) server. User-friendly browsing, searching facility is integrated for easy data retrieval. It also includes useful tools like BLAST CrisprGE, BLAST NTdb and CRISPR Mapper. Considering potential utilities of CRISPR in the vast area of biology and therapeutics, we foresee this platform as an assistance to accelerate research in the burgeoning field of genome engineering.
Database URL: http://crdd.osdd.net/servers/crisprge/.

Dieser Aufsatz gibt eine Übersicht über die aktuelle Deba􏰁e zu den neuen Me- thoden der Genomedi􏰀erung und legt dar, welche Bedeutung der öffentliche Diskurs dabei hat und wie er aktuell geführt wird. Dafür entwickeln die Autoren ein Forschungsdesign, um die Effekte unterschiedlicher Vermi􏰁lungs- und Par- 􏰀zipa􏰀onsformate auf den Dialog zwischen Wissenscha􏰂 und Öffentlichkeit zu untersuchen. Die verschiedenen Formate werden am Museum für Naturkunde Berlin getestet. Das Museum ist mit über 800.000 Besuchern im Jahr 2016 das besuchsstärkste Naturkundemuseen in Deutschland und hat sich zur Aufgabe gesetzt, den Dialog zwischen Wissenscha􏰂, Poli􏰀k und Öffentlichkeiten zu in- tensivieren. Dies ist für die neuen Methoden der Genomedi􏰀erung besonders wich􏰀g, weil Entscheidungen über ihre Regulierung und ihren Einsatz weitrei- chende Folgen haben – sowohl ökologische als auch kulturelle im Hinblick auf das Verhältnis zwischen Mensch und Natur.

This paper gives an introduction to the inter-disciplinary special section. Against the historical and ethical background of reproductive technologies, it explores future scenarios of human reproduction and analyzes ways of mutual engagement between fictional and academic endeavors. The underlying idea is that we can make use of human reproduction scenarios in at least two ways: we can use them to critique technologies by imagining terrible consequences for humanity but also to defend positions that favor scientific and technological development.

Genome editing with CRISPR/Cas has rapidly gained popularity. Base editing, a new CRISPR/Cas-based approach, can precisely convert one nucleotide to another in DNA or RNA without inducing a double-strand DNA break (DSB). A combination of catalytically impaired nuclease variants with different deaminases has yielded diverse base-editing platforms that aim to address the key limitations such as specificity, protospacer adjacent motif (PAM) compatibility, editing window length, bystander editing, and sequence context preference. Because new base editors significantly reduce unintended editing in the genome, they hold great promise for treating genetic diseases and for developing superior agricultural crops. We review here the development of various base editors, assess their technical advantages and limitations, and discuss their broad applications in basic research, medicine, and agriculture.

Changes in Earth's climate at the end of the last ice age brought about seasonal conditions that favoured the cultivation of annual plants like wild cereals, helping to launch the agricultural revolution. Earth's climate is changing again, mainly through the effects of human actions on the biosphere. To feed a projected population of 9.6 billion people by 2050 while reducing agriculture's carbon, nitrogen, and environmental footprints requires a revolution in food crop productivity and a deeper understanding of the interplay between sustainable food production and natural ecosystems. These goals cannot be achieved without making appropriate use of advanced technologies. Genome-wide association studies, marker-assisted selection, and genomic selection of orphan crops in developing countries can help enhance yields, nutrition, disease resistance, and crop resilience in the face of climate change. With major cereal crop yields stagnating or in decline, successful C4 photosynthesis engineering of rice and wheat and nitrogen fixation engineering of rice, wheat, and maize would have enormous consequences for crop productivity, environmental remediation, and land, soil, and water conservation. Next-generation DNA sequencing, genome editing, synthetic biology, and molecular modeling provide the tools needed for these ambitious efforts to succeed. Innovative food crop bioscience and healthy ecosystems constitute a symbiosis for the Anthropocene.