Tools like CRISPR have revolutionized the way we look at personalized medicine, by manipulating DNA, and opening up possibilities for a cure that didn’t exist until now.
Vijay Elango, our next pathbreaker, works as Research Associate at CRISPR Therapeutics (Cambridge, Massachusetts), a company that develops therapeutic drug products using engineered immune cells derived from patients.
Vijay talks to Shyam Krishnamurthy from The Interview Portal about Gene Therapy, an emerging technology that has the potential to treat serious genetic diseases with precision, safety and efficiency.
For students, a career in cutting edge technologies is as unpredictable as it gets. But tough moments such as these show you where your true interests lie !
Vijay, can you tell us about your growing up years?
I grew up in Salem (Tamilnadu), surrounded by a family environment where life sciences drew curiosity and attention. My grandfathers were both agricultural biologists, my father an anesthesiologist, my mother a nutritionist and my immediate uncles were physicians.
Growing up (1990s) before the birth of Google, it was books and conversations that filled up our time, stimulating our interests in general knowledge or ‘GK’ as we called it. In school, other than biology, I always enjoyed English, History and Geography. I was fairly active with participation in quizzes, volunteering activities involving clean-up drives and speech competitions. Music was a major contributor that influenced my thought process during school days. I was lucky to have the chance to learn both vocal carnatic music and classical piano and had great teachers with whom I still chat, reflecting on music and beyond.
Until higher secondary, though I was not fixated on any specific field as a choice of career, I knew I wanted to stay within the STEM domain. Further along, scoring quite well in grade 10 biology gave me the illusion that I would become a doctor (cardiologist). So, I rode that thought and wanted to pursue medicine after school, although my father wasn’t very fond of my choice.
What did you study?
Unable to ride the ‘medicine’ bandwagon, I enrolled in a Bachelor of Technology program specializing in Biotechnology for my undergraduate studies. Being in a 4 year interdisciplinary program in an engineering college, I felt that not enough justice was done in terms of practical exposure to topics involving Genetics, molecular biology, cell biology given the limited access to infrastructure. So, towards the end of my bachelor’s degree, I really wanted to see myself inside labs, getting that practical exposure and research training in topics like genetics. In 2008, I chose a one year Master’s by research program at the University of Glasgow, with a focus on Biomedical Sciences.
What made you choose such an offbeat, unconven tional and uncommon career?
I could think of a couple of events that really made me choose this path. I think around the same time as the Y2K problem (year 2000) was the news about the completion of the ‘rough draft’ of the human genome through the human genome project. It was like a science fiction story.
I had no clue what a genome would look like, nor did I fully understand its functional significance. My exposure was limited to just mitosis and meiosis. In 2007, during summer, I had the opportunity to spend a month at the Department of Cytogenetics, Cancer Institute – Adyar, where for the first time I visually saw images of the chromosomes. And through techniques like ‘FIsH’ , seeing the chromosome lit up with so many colors (computer generated) was really inspiring. We all love visuals and so did I, a visual that was the turning point in influencing me to learn more about DNA.
My second turning point involved a pious moment, literally ! By 2007 /08 I was getting familiar with the idea of ‘cloning’, ‘cutting DNA strands’ and ‘joining cut DNA strands’. At the temple, seeing a circular garland on the altar, with patches of flowers of different colours, aligned in a sequence gave me a visual of a ‘recombinant plasmid’ or the idea of ‘cloning’ a foreign ‘gene’ into a host backbone. It might sound silly, but in this instance it would be fitting to refer to HIM as one my key influencers.
Subjective feelings aside, I was also influenced in school by the narratives of Richard Feynman (although I was mediocre at Physics) and Francis Crick’s narrative about discovering the structure of DNA in his book, ‘ The Double Helix’. I was also lucky to have a couple of inspiring teachers in college who taught us to visualize Cell biology and Genetics.
How did you plan the steps to get into the career you wanted? Or how did you make a transition to a new ca reer? Tell us about your career path
For my undergraduate project, I worked at the Madras University, Taramani on a topic related to mitochondrial DNA. That was my first hands-on experience working independently on a project making me realize learning science through books and doing science in a lab were two worlds apart. Failed experiments, knowledge gaps, long hours in the lab, poor time management and silly experimental mistakes really exposed areas where I could improve if I wanted a career as a researcher. My one year lab intensive Master’s program was a painful yet an excellent learning phase where I was simply thrown out into the ocean and told to swim. It taught me a lot and since then I grew stronger and stronger exploring myself and my interests working on different projects. I returned back to India with sufficient training in a research lab and wanted to work at a place like NCBS on research projects before deciding on PhD topics to study.
Instead, I had my first internship (unpaid) at IIT Madras working on a project for a startup pharmaceutical company. The project seemed exciting and I was really eager to work with imaging / fluorescent confocal microscopes to visualize cells undergoing drug treatments. However, due to administrative restructuring I was instead working on another instrument called the HPLC, profiling drug compounds and in a month’s time I was bored with the monotony, let alone not being paid.
But being physically present in IIT Madras, I was able to interact with other lab members, visit their labs and know more about the spectrum of research projects available. Through the IITs, I came to know about the recent establishment of IISERs ( Science education institutes spread across India) and found interest in a couple of labs, working on a topic similar to what I had pursued in my Masters in the UK. I applied, went down to the lab in Pune, was interviewed by the team there and was offered a position as a project assistant. My first paid job. It was a great feeling. Though the area of research was very similar, I was now working with ‘fruit fly’ as a model organism. Fruit flies are tiny, social and usually red eyed fruit loving flies with their genome architecture almost a 70% match to that of humans. I was not a ‘fruit fly’ biologist and hence it was a steep learning curve to know more about the genetic manipulations that can be done on these flies. Research labs work with fruit flies because they are excellent organisms to study genetic phenomena. By building on concepts like Mendelian genetics you can simply engineer their genes. For example, we can generate yellow eyed flies, flies with more stubble, bigger wings or simply just flies of one gender !
Spending a couple of years in the fly lab and being interested in them as model organisms, I was ready to pursue a PhD. Through my lab members who had recently been to an international ‘Fly biology’ conference in Taiwan, we learnt that there were some really exciting research groups in the Far East ( Eg. Japan, Taiwan), welcoming international students for PhD studies. I loved the idea of exploring the east for travel and pursuing a PhD there, so I readily took the offer.
My time in Taiwan was truly inspiring both in terms of lab and life experiences. Research culture in my lab was top notch and competitive. Continuing my PhD research using fruit flies, I was working on problems related to the peripheral nervous system, in particular structural architecture of dendrites (tree branch-like structures of a neuron) and how they are affected during development. Experiments would involve extensive ‘photography’ of these neurons using a specialized instrument called the confocal microscope. Fruit flies are genetically manipulated to have specific protein targets in neurons ‘tagged’ with another fluorescent protein to track its location, structure, and study its function during development. All these nanoscale structures can be viewed under a confocal microscope, where a laser beam excites these ‘tags’ and fluorescences the neuronal structure (Image attached)
Image: A fluorescently illuminated neuron with branched dendrites viewed under a confocal microscope.
From being lost in translation to making some truly life long friends, Taiwan was a magical experience. Until today, the one thing I miss when I reflect back on my career path, is not completing my PhD. Although I took a break from my PhD in Taiwan due to health reasons, I took up a temporary research position for a year, closer to home at NCBS, Bangalore working for the first time on the technique called CRISPR.
CRISPR technology is a DNA editing tool adapted from prokaryotes, mainly bacteria. As an acquired immune response against viruses that attack them, bacteria harbour viral ‘CRISPR’ sequences in their genome to mount a defence against future viral attacks. This way, a CRISPR sequence in partnership with an associated bacterial protein called Cas9 would ‘bind’ an infecting virus DNA and then let the protein cut the ‘foreign viral DNA’.
Realizing the potential of this mechanism for editing DNA, several customizations were made, with many components of this mechanism adapted for use to edit mammalian cells (like human cells). The central dogma of biology goes like DNA makes RNA, RNA makes protein. Typically, pharmaceutical drug products involve treating diseases at the protein level. What about treating genetic diseases where faults lie at the DNA level ? Here, tools like CRISPR can help treat diseases at the DNA level, acting as a ‘pair of scissors’ to cut and edit DNA.
Towards the completion of my 1 year contract at NCBS/ inSTEM and having already drifted away emotionally from my PhD work, my chances of returning to Taiwan were almost nil.
In NCBS, both my mentors were Japanese and almost all the faculty in Taiwan and NCBS had training in the US, inspiring me to work there. Besides, I wasn’t really keen on staying back in India for a long term career in life sciences research. So I started looking at options to move to North America to start afresh. Canada at that time initiated a very friendly immigration policy and that’s how I ended up in Toronto in 2016 to restart my career in life sciences research.
Finding a desirable job in Toronto was a challenge and I did not want to settle for one outside my field of interest. So given the choice between research and money, I worriedly took up a research position in a university. Although I had a frugal lifestyle, I spent three great years in a lab working on regenerative medicine and gained valuable teaching experience as well. Given the current scenario of organ shortage for transplant, imagine re-programming a non-cardiac cell to become a cardiac cell to treat a diseased condition. That’s the kind of work I was doing, looking at factors that affect the re-programming of a fibroblast cell into a cardiac (heart) cell. Reprogramming, in simple words, refers to tweaking the genome, instructing them to stop expressing non-cardiac genes and express more cardiac-related genes or ‘behave’ like a cardiac cell. A part of the approach also includes screening for hundreds of nuclear proteins ( DNA interacting proteins: Imagine them as fingers touching the piano to make music) that would facilitate re-programming. These screens are facilitated using bioinformatic tools that help categorize and prioritize these proteins as likely candidates. Typical of a research lab scenario, funding constraints started directly affecting my remuneration and given the cost of living in Toronto, after three years, I decided to move to the industry for a job.
Research in academia is like an off-road driving experience, while that in an industry is typical of driving in an expressway. The former tests your perseverance, originality and ability to maneuver, while the latter tests your consistency, discipline and compliance to the system and your co-drivers. When I moved to the industry, I took up a position as a research analyst, translating the molecular biology skills that I learnt earlier towards routine testing of nutraceutical products using techniques such as a ddPCR, ELISA and qPCR, screening for bacterial genes and proteins. This is the work that takes place before you see those ‘non-GMO’ certified labels on the sides of your favourite protein powder pack. Industry teaches you to be meticulous in documentation, adhere to criteria defined to qualify/pass an assay and collaborate with cross-functional review teams before you release your results (certification of analysis). Working in a Contract Research Organization (CRO) for multiple clients, I was exposed to other projects as well. Of particular mention was my role to generate a genetically modified strain of the Cannabis plant (cannabis sativa). This was an exciting yet challenging project because I had no experience with plant biology but had the opportunity to use CRISPR on plants. Although the project was shelved after a year, it was a great learning experience knowing more about plant physiology, health, their growth curve, nutrient requirements etc. Towards the end of the project I started having genuine interests in horticulture, learning to care better for plants.
Presently I work in Boston, in the analytical development team of a therapeutics company that focuses on gene therapy based medicines. My work experience in the CRO environment helped me get this position here in Boston.
How did you get your first break?
I would consider the opportunity as a research assistant at IISER, Pune as my first break, because I was able to get back to research work and following which I was able to build my network/ contacts and move forward.
What were some of the challenges you faced? How did you address them?
No road is smooth and there are always challenges along the path. Two challenges that really tested me other than the time spent to fill knowledge gaps were :
Challenge 1 : Funding
Research labs depend on external sources, mostly government agencies for funds and the outcome of the allotment directly affects lab function and access to lab resources. There have been times involving pay cuts, axing of projects, budgeting of resources and falling back to more laborious manual lab techniques just to save costs. The only way I addressed this challenge was by looking at the big picture and keep at it during tough times, because the belief was that this phase would get better.
Challenge 2 : Health
Another personal attribute we tend to oversee is our mental and physical well being. Unfortunately, I suffered a couple of health related issues for a few years which slowed down the pace of my work and brought down my mental strength. I was close to switching fields due to health reasons, but eventually managed to survive in this field through challenging times. I would attribute my faith in a higher power as one of the reasons for my endurance.
Where do you work now?
I currently work at CRISPR therapeutics, Boston as a research associate in the analytical development team
What problems do you solve as Research Biotechnologist?
The company focuses on developing therapeutic drug products using engineered immune cells derived from patients. Engineering these immune cells involves genetically manipulating the existing cell using gene editing techniques such as CRISPR and adding the ‘healthy’ gene using ‘useful’ viruses called AAV. These engineered cells put back into the patients help treat genetic diseases including cancer. As members in the analytical development team, we monitor the critical attributes of various components that go into making the drug product. For eg, the purity of the introduced viruses, specificity of the genetic edit, efficiency of the engineered cell to treat the disease etc. This analysis would require prior working knowledge of the biology involved, some lab skills and hand-eye co ordination to handle samples and techniques, time management, ability to troubleshoot if there are deviations in the results. These skills come through years of practice and interest.
What’s a typical day like?
A typical day involves a combination of lab work, data analysis (desk work), lab meetings involving various other groups and ppt presentations if any. And this is some thing I love about my job, where I am both mentally and physically busy.
How does your work benefit society?
Gene therapy is an emerging technology that has the potential to treat serious genetic diseases with precision, safety and efficiency. Tools like CRISPR have revolutionized the way we look at personalized medicine, where diseases can be treated specifically at the required nucleotide bases (DNA structure) of an individual, opening up possibilities for a cure that didn’t exist until now.
Tell us an example of a specific memorable work you did that is very close to you!
If I could think of a proud moment, it would involve my work at NCBS, Bangalore where I independently handled the CRISPR technique for the first time and was able to demonstrate the expected results to my Japanese collaborators. There were moments during the project when one of my mentors believed that the approach wouldn’t work, but I was insistent that it would work and we marched ahead with the plan. Gladly, one year of hard work paid off and it was a proud moment when the experiment worked.
Your advice to students based on your experience?
Have a bit of faith during tough times, explore, take time to enjoy life outside work to learn from the world around. Stay curious and don’t forget to take care of your mental and physical health.
A future in the gene therapy space.