Recommendation Letter for National Interest Waiver (NIW)

RE: Form I-140 Petition of EB2 NIW (National Interest Waiver) for Dr. Mukesh Maharjan

To whom it may concern;

It is a true pleasure to write this recommendation letter for the EB2 NIW (National Interest Waiver
petition of Dr. Mukesh Maharjan who is currently working as a Postdoctoral Associate at the
University of MD Anderson Cancer Center, Houston. He is, without question, one of the most
productive researchers I have ever known.

As for my qualifications to provide this evaluation, I am ________ (Ph.D. in Biochemistry and Molecular
Biology). I received my Bachelor of Science from __________. I received my Ph.D. in Biochemistry
and Molecular Biology from the University of ___________, in _________. In __, I joined the faculty of
the University of _________, where I stayed until _______, before joining the University of _______ as a
professor of Biochemistry and Molecular Biology. I have over _______ publications in the scientific
literature and hold over 22 US patents. I am a member of the National Academy of Sciences. Some of my
notable awards include: ACS Award in _________ Biochemistry and Molecular Biology (Year),
________ Section (____) Distinguished Speaker Award (Year), ACS A. C. Cope Scholar Award (1994),
____ Society for the Promotion of Science (JSPS) Fellowship (1995), American Academy of Arts and
Sciences Fellow, (1996),  ………….

Being in close contact with Dr. Mukesh Maharjan, and discussing various scientific projects with him, I
quickly realized how experienced and innovative he is. Up to now, he has conducted numerous and
completely different projects.  I can confidently assert that running each of them very well does indeed
require a unique capability which in my expert opinion is called true research capability, for which years
of industrial and academic activities have to be taken, for which energy and time have to be devoted, and,
most importantly, for which creation and innovation have to be offered. These are the unique features that
Dr. Mukesh Maharjan possesses, distinguishing him from his other peers.

Dr. Mukesh Maharjan research area can be divided into four distinct yet interconnected areas of soft
matter as follows.

1. ________________________
2. ________________________
3. ________________________
4. _______________________

As his Ph.D. project, Dr. Mukesh Maharjan designed as Research Associate at Louisiana State
University (LSU), Department of Biological Science. During his Ph.D., he gained invaluable
knowledge and experience as below:
 i. Explored insulator protein BEAF and its protein partners to elucidate the role of it in maintaining
the chromatin dynamics in Drosophila.
ii. By performing experiments like Co-IP, Protein expression (in Drosophila, insect cell lines & E.
Coli), purification (from embryo, insect cell culture and E. Coli) & identification (by mass
spectrometry & data analysis) Yeast-2-hybrid, rough eye phenotype assay, scanning electron
microscopy (SEM) discovered a list of candidate proteins that were not previously identified in
insulator functioning of BEAF.
iii. Established modified versions of PCR like TAIL PCR and Splinkerette PCR to successfully map
the randomly integrated target DNA into the genome. Designing specific primers using Primer3
and BLAST.
iv. Established an in-vivo transgenic animal model suitable for enhancer trapping system by creating
drosophilized stable transgenic line using phiC31 Disintegrase system for the first time in its
field.
v. Performed transient transfection assays with luciferase gene reporter assays to test the promoter
activity of candidate Drosophila genomic sequences in KC and S2 cell lines. • Instructed 5
researchers (including 2 graduate students) and completed 3 projects with 3 scientific research
paper sin peer-reviewed journals.
vi. Guided and taught classes of >180 undergraduate students for more than 3 years with satisfaction
rate of > 98% form the students.
vii. Prepare up to date lecture slides and exams to guide students and simplify complex genetics,
biochemistry and molecular biology topics.

viii. Manage the lab logistics, organize lab chores, and create SOPs, documentation and budgeting of
lab expenditures for efficient running of the lab.

Next he continued his researches in Baylor College of medicine, Houston, and worked and gained
experience as below:

i. Lead a project by designing and planning experiments that involve bone marrow transplant, Flow
cytometry, behavioral testing, primary cell culture, ddPCR and qPCR to elucidate the links
between clonal hematopoiesis and environmental stress.
ii. Analysis of low gene copy numbers by ddPCR and detecting extremely low copy DNA from
mycobacterium infected blood by qPCR.
iii. Established and optimized T cell and macrophage co-culture with primary microglia culture to
evaluate inflammation related activation of microglia.
iv. My overall goal is to elucidate the links between hematopoietic cell function and
neurodegenerative disease
After that, he continued his Post-Doctoral Associate in MD Anderson Cancer Center, Houston, and
worked and gained experience as below:

Explored insulator protein BEAF and its protein partners to elucidate the role of it in maintaining the
chromatin dynamics in Drosophila.

i. By performing experiments like Co-IP, Protein expression (in Drosophila, insect cell lines & E.
Coli), purification (from embryo, insect cell culture and E. Coli) & identification (by mass
spectrometry & data analysis) Yeast-2-hybrid, rough eye phenotype assay, scanning electron
microscopy (SEM) discovered a list of candidate proteins that were not previously identified in
insulator functioning of BEAF.
ii. Established modified versions of PCR like TAIL PCR and Splinkerette PCR to successfully map
the randomly integrated target DNA into the genome. Designing specific primers using Primer3
and BLAST.
iii. Established an in-vivo transgenic animal model suitable for enhancer trapping system by creating
drosophilized stable transgenic line using phiC31 Disintegrase system for the first time in its
field.
iv. Performed transient transfection assays with luciferase gene reporter assays to test the promoter
activity of candidate Drosophila genomic sequences in KC and S2 cell lines.
v. Instructed 5 researchers (including 2 graduate students) and completed 3 projects with 3 scientific
research paper sin peer-reviewed journals.
vi. Guided and taught classes of >180 undergraduate students for more than 3 years with satisfaction
rate of > 98% form the students.
vii. Prepare up to date lecture slides and exams to guide students and simplify complex genetics,
biochemistry and molecular biology topics.
viii. Manage the lab logistics, organize lab chores, and create SOPs, documentation and budgeting of
lab expenditures for efficient running of the lab.

While he was Bsc in Biotechnology designed Research Associate at SANN International College,
Purbanchal University, Nepal he gained invaluable knowledge and experience as below:

i. Developed virus-free plants by performing ELISA, apical meristem culture and greenhouse plant
breeding. Distributed virus-free plants to local farmers for local agriculture.
ii. Studied molecular characteristics of rice varieties of different altitudes of Nepal and successfully
categorized them according to their seed dormancy.
iii. Received Scholarship for 6 semesters (out of 8) of free university and tuition fee by being the top
undergraduate student in the biotechnology department in the university.

While pursing Bsc in Biotechnology, he worked as Research Technician. Please find some of his
Research Technician work:
 i. Production of virus free plants (Potato, cardamom, orchid etc.) through apical meristem culture
for local farmers, also generating revenue to sustainably operate the plan t molecular biology
laboratory.
ii. Conducted tests to diagnose plant virus related diseases with the DAS-ELISA techniques.

Next he continued his researches in Nepal Agricultural Research Council (NARC), Khumaltar, Nepal
and worked and gained experience as below:

i. Acted as a lead scientist to study the marketplace and built connections with vendors and
ultimately established a brand-new molecular biology lab to study seed biology for the
improvement of current seed technology in Nepal.
ii. Mentored 3 associates and studied 41 Rice varieties of different altitudes of Nepal by
troubleshooting problems related to ELISA, primer design and PCR that led to the successful
completion of the project 2 months ahead of the deadline.
iii. Studied the marketplace and built connections with vendors that finally resulted in establishing a
brand-new full functioning Molecular-Biology Lab in the Department.

In sum, I am absolutely convinced that Dr. Mukesh Maharjan will be very productive in his future
research activities, as he has been, and that he will greatly contribute to the national interests of the USA
Therefore, I fully support and recommend his application.

Sincerely yours,

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Professor (Name)
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(University Name)
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