First person – Jacob Odell Free

Jacob Odell

How would you explain the main findings of your paper in lay terms?

Biological researchers often use ‘tags’ to label a protein of interest so that we can observe where a protein goes within the cell and how it behaves in response to various stimuli. For example, tags have been used to investigate lamins, which are intermediate filament proteins that provide structural support to the nucleus and are essential for normal function of mechanically active tissues, such as muscle. Although tags are vital for studying protein function, in some cases they might compromise the normal function of the protein to which they are attached. In our study, we investigated the effect of different tags on the function of lamin A using a variety of biophysical, biochemical and microscopy-driven approaches. Surprisingly, we found that tags impaired some functions of lamin A but did not affect others. Tagged lamins performed equally well as non-tagged controls in experiments designed to evaluate nuclear morphology and nuclear envelope integrity, but tagged lamins were defective in their proper localization, ability to prevent nuclear deformation and ability to interact with the integral nuclear membrane protein emerin. Our data serve as a reminder to the cell biology community that tags should be used with caution, and that in many cases, tags might impact the function of a protein of interest.

Were there any specific challenges associated with this project? If so, how did you overcome them?

One of the big challenges associated with this project was implementing a system that allowed for a fair comparison between differently tagged lamin A constructs. We needed to express different constructs at similar levels, because the degree of lamin A expression can directly impact the output of some of the assays we used to measure the function of the lamin protein. Initially, I used a lentiviral expression system with a constitutive promoter to drive lamin A expression, but I was never able to achieve precise control and could not match the expression levels between the different constructs. This led us to pursue the doxycycline-inducible system used in the paper, with which we can precisely control protein expression by titrating the levels of the inducer compound doxycycline in clonally derived cell lines. This approach reduced the heterogeneity in lamin A expression between cells and allowed for us to more cleanly compare the effect of different tags on lamin A function in a way that would likely not have been possible using previous techniques.

When doing the research, did you have a particular result or ‘eureka’ moment that has stuck with you?

One of the first things I did once I generated the cell lines expressing the differently tagged constructs was to stain the cells for lamin A and look at them under the microscope. I remember initially being excited that the staining worked, and then getting even more excited that I could already see differences between cells expressing the tagged and untagged constructs by eye – the tagged lamins were enriched in the nucleoplasm, whereas the untagged lamin was enriched only at the nuclear periphery. To me, microscopy data is the most intuitive type of data because I can see for myself what is going on, and differences between conditions or cell lines immediately become apparent. That first glimpse of a striking difference between the different constructs stuck with me as I was working on this project and made me excited to work on the subsequent experiments.

Why did you choose Journal of Cell Science for your paper?

I thought our paper was an excellent fit for the Short Report format offered at Journal of Cell Science because our story is straightforward, relatively compact and self-contained, and will hopefully be useful for the broader scientific community. I found the review process to be very reasonable and helpful, so I would definitely consider submitting future work here as well.

Have you had any significant mentors who have helped you beyond supervision in the lab? How was their guidance special?

One person who has significantly influenced my scientific journey is Dr Andrew Scala, who I met while I was a student at Dutchess Community College. Given that the college did not have an undergraduate research program, I did not have any real research experiences until Dr Scala noticed my interest and effort in the genetics lab course he taught. We ended up talking about careers in science and ways to get involved in hands-on research. Based on his recommendation (and also because he drove me 2 h to attend an open house after class one day!), I was able to participate in a Research Experience for Undergrads (REU) program hosted at the New York Department of Health. From this experience, I learned that I loved doing bench work and had a knack for research, and the REU also opened up many doors for me, including being able to pursue a PhD here at Cornell University. I am eternally grateful that Dr Scala took an interest in my passions and gave me a chance to develop into the scientist I am today.

What motivated you to pursue a career in science, and what have been the most interesting moments on the path that led you to where you are now?

One of the things that motivated me to pursue a career in science was a conversation I had during my first research project with my PI, Dr Michael Koonce. While asking me questions about my results and my interpretation of an experiment, he said that I was now the world expert on this very specific assay using these cells with our particular setup. This left an impact on me because I thought it was so cool to be the first person to ever do this specific experiment and to be the first to learn something completely new about how the world works. I have always wanted to understand more about why things are the way they are, and I think that this feeling, which is something that cannot be found in many other professions, motivates many scientists.

What's next for you?

I am planning to wrap up my graduate work in the next 6 to 9 months, and I am starting to actively apply for both postdoc and industry positions.

Tell us something interesting about yourself that wouldn't be on your CV

I am a big fan of collegiate ice hockey and my fiancé and I went to every Cornell hockey home game last season. We have season tickets for this year as well! Go Big Red!