Author Archives: Caitlin Lamb

The end of the chapter

At the beginning of the program, I made some goals, and fortunately, I reached those goals.  As a refresher, here were my goals I wanted to accomplish:

  1.  I would like to learn to make mistakes.
  2.  I would like to learn to be patient.
  3.  I hope to be able to communicate and learn from the community around me.

The first and second goal go hand-in-hand.   I did make some mistakes during the program.  I did not use the correct amount of DNA, I made a mistake during a presentation (i.e. mixed up some background information), produced blots where the positive control failed ( I’m not sure what went wrong), and produced blots where the IPs did not show up, when the inputs did (again, I am not sure what went wrong).  However, I’ve learned to troubleshoot, and I am continuing to learn to troubleshoot and accept that things do not always work and you have to keep trying.  On the same note, cell cultures (especially when starting with single cells) grow very slowly, western blots with IPs takes approximately 4 days, so you need to be patient.  If the science does not work, you will have to start again taking even more time, thus, science takes patience and is all about delayed gratification.

The third goal was also achieved.  Through BSURF led by Dr. G and Jason, my peers and I were able to listen to amazing talks, give chalk talks, and will give a poster tomorrow. Through my lab, I was able to present what I have accomplished in the summer, and learn from others in my lab.  I feel very lucky to have been able to communicate about my science and learn from others as well.

I am really thankful for this experience, and I truly feel that this experience allowed me to see what it is like to do research full time. The BSURF program makes me feel excited about the possibilities of attending graduate school but continues to leave the next step beyond graduate school open.  What do I want to do?  Do I want to go into industry? Do I want to try to start a startup? Do I want to go into academia?  What are my options?

I am excited to continue my work in this lab, to build strong relationships, and to learn more about my next steps.

Thanks for experiencing this program with me!

”So long, farewell…goodbye

This was the final blog post.

The Penultimate Blog-Journeys of scientists

Over the course of the program, my fellow BSURFers and I had the opportunity to listen to various scientists talk about their research and their journey.

All scientists had something amazing, thoughtful, and encouraging to say about their science, and to someone who may consider pursuing science.  One talk by Dr. Lawrence David really stood out to me.

First, he has interesting research by looking at the human microbiome, asking for, and handling human fecal samples.  He also shared ongoing work with prebiotics and the microbiome and even informed us that he is still taking participants.

What stood out was not only his research but how he explained his journey through a series of questions that he asked himself. I enjoyed the stories of not knowing what he wanted to do but he had an it wouldn’t hurt to try attitude. I liked his anecdote of saying graduate school was an amazing experience, and he was even able to get his PI to fund his trip to Asia to eat nothing but street food (and collect and look at his fecal samples) for two months.  Additionally, I found it interesting that he joined new labs (where the PI was within his first year) all throughout his career. I thought it was great how he says even as a professor he doesn’t know what he wants to do, mentioning start-ups, mentioning if it was right not to do med. school, but always follow up with how he enjoys and loves what he does.

Finally, the thing that I thought was interesting was how he became interested in science: Jurassic Park. He mentions how a great advocate for science could also be Hollywood, which I found interesting.

That’s it… on next weeks blog I will wrap up my BSURF experience.

 

That’s Life…science

A song made famous by Frank Sinatra, That’s Life, talks about some important facts/lessons about life that also connect to science.

The song talks says life is all about the highs and lows about life.  Science, as I found out, is also all about the highs… and the lows Things do not always go according to plan.  The deletion that you wanted to make, does not delete.  Sometimes, you may try a mutagenesis reaction multiple times with no luck, but that is okay.  Science takes time, and eventually, it will come back around and you may get a result, so don’t quit.

Over the summer, I attempted to delete Sec24D from SW1353 (a chrondrosarcoma cell line), and SAOS-2 (a osteosarcoma cell line) with no luck. Additionally, I’ve attempted several mutagenesis reactions with minimal successes.  These were the lows.  However, I did have some highs. Other projects and experiments went smoothly, and I am happy for all experiences.

I am also thankful for my successes, and “failures”.   There is more to say about what I did over the summer, but I would like to save it for my poster presentation that will be held on July 27, 2018, and future blogs.

As a teaser for the poster presentation, I will post a draft (science is all about sharing current knowledge about a specific topic, so please understand this is subject to change) of my project’s abstract.

 

What role does SEC24D O-GlcNAcylation play in collagen secretion?

The COPII complex mediates traffic from the endoplasmic reticulum (ER) with five major components: SAR1, SEC13, SEC23, SEC24, and SEC31. Posttranslational modification plays an important role in the regulation of the COPII complex. Our lab and others have found that O-linked β-N-acetylglucosamine (O-GlcNAc), a single sugar modification added to serine and threonine of intracellular proteins, decorates many COPII components. I mainly focused on O-GlcNAcylation of SEC24D, a subunit of the COPII complex, because in humans, SEC24D mutations cause a subtype of osteogenesis imperfecta, a collagen trafficking disorder. I wanted to answer the question, “What role does SEC24D O-GlcNAcylation play in collagen secretion?” I hypothesized that SEC 24D O-GlcNAcylation is necessary for normal collagen secretion. To test this hypothesis, I am deleting SEC24D in osteosarcoma cells using CRISPR-CAS9 and creating unglycosylatable SEC24D mutants to reintroduce into the SEC 24D deleted cell line.  After reintroducing the mutants, I will look at collagen secretion using immunofluorescence microscopy (IF) to determine the role of SEC24D O-GlcNAcylation in collagen secretion. We expect the IF to show more collagen in the ER of the mutants than the wild type, showing SEC24D O-GlcNAcylation is important in collagen secretion.

 

Life in the lab

For me, there are things that I do on a daily basis, but there are also things that I do that depend on how a previous day’s work turned out.

Most days, I arrive at the lab I check in with my mentor Brittany and plan out the day’s work.  Then we usually check on our cells to see if we need to split them, how to split them (1:10, 1:20, etcetera), or if the cells just need a media change.

Once we make a plan I get to work. On a given day, I might be making gels, and running gels, to get ready to develop a western blot the next day.  I might also be collecting cells, and lysing the cells to either do an Immunoprecipitation (IP) or a normal western blot.  Additionally, I will sometimes treat the cells, or transfect the cells in order to delete a certain gene or express a construct so that I can see what is happening to expression levels when I treat my cells.  Finally, I may also step away from the cells and attempt to do a mutagenesis reaction.  In order to do this, I will first set up the mutagenesis reaction (it is similar to a PCR reaction but with a different primer), and run the mutagenesis reaction.  The next day (day 2), I will do a DPN1 to get rid of the original template (so that you have your mutated DNA).  Then I will transform the new DNA into bacteria, and plate the bacteria.  The next day(day 3), I will collect various colonies and grow them out.  Then on day 4, I will isolate the DNA from the bacteria and send it off for sequencing.  On day 5, I will receive the sequence and go from there.

Some days, the science does not go as you hoped it will and some experiments have to be done again, some might have to be changed, or you might have to stop the experiment altogether.  All in all, the days in the lab in my experience is slightly different each day and builds upon the previous day.  Each day, I learn something new and it has been a great experience.  I am learning to be patient and realize it is okay if things don’t go as well as you hoped… more on this on next weeks blog where I will talk about the highs and lows of my project.

 

 

Everyone gave an amazing chalk talk

This week, I had the opportunity to listen to everyone give a chalk talk about their project.  It was nice to understand details about their project,  such as Alzheimer’s, and DMD instead of thinking, okay he works in an x,y,z lab.  Additionally, I enjoyed how everyone was able to present such complex topics in a short amount of time in a way that was easy to follow.

As someone who attended the NC Zoo school, I had the opportunity to go into the zoo on a daily basis.  If time allowed me to, I enjoyed going to the African side of the NC Zoo to watch the chimpanzees and baboons.  When Christine presented the idea that both the highest ranking (alpha) and lowest ranking baboon experiences the same amount of stress, but it might be due to a different kind of stress I was all for it.  Over the summer she is planning to answer the question, “Do alpha baboons and low ranking baboons experience different stressors?”.  She is planning to look at energetic stressors, so she will analyze a thyroid hormone(1), T3, in a wide range of baboon fecal samples. Another thing that caught my attention about the presentation, was the fact that there was a significant decrease in stress from the alpha to the second rank baboon (beta).  Finally, it was great to learn the behind the scenes of this research project.  She talked about how they have an area in Kenya and scientists in Kenya watching the baboon’s behavior (looking for changes in rank), collecting the samples, and shipping them back to Duke for analysis.

This is an amazing project and I can not wait to see the outcome of this project and other projects on July 27, during the poster presentation.

Next week, I will provide a glimpse into my daily routine (A day in the life).

1-thyroid hormones allow you to indirectly measure energetic stress because it is based on changes in metabolism.

An interview with Dr. Michael Boyce

Dr. Boyce has enjoyed science since as early as elementary school, however, he was not sure about a career in science until later on in his life.  As an undergraduate student at Harvard College, he saw the biochemistry major and thought the courses were interesting.  In addition to courses required for the biochemistry major, Dr. Boyce found another interest, art history.  At this point, he realized he liked both art history and biochemistry and he asked himself, “Do I want to stick with biochemistry, or do I want to switch to art history.” To answer this, he pictured himself in both situations, as an art historian, and as a scientist.  He did see himself as a scientist, but he could not picture himself as an art historian.  He joined a lab at Harvard, and decided to get a PhD. with the idea of , “if this does not work out, I will pursue something else”.  Therefore, he was able to go to Harvard Medical School for his PhD. in molecular biology, he enjoyed graduate school, made great connections with students and his Principal Investigator (he even stays connected with them to this day using social media!).  With this experience, he decided that having his own lab is something he might enjoy, so he decided to do a post-doc with the same mentality of, “if this does not workout, I can do something else”.  Therefore, he went to UC Berkeley to do a post-doc in a Chemistry lab.  From the differences of research topics, he met many great friends, and learned new techniques.  Finally, he applied to faculty positions, where he is now an Assistant Professor of Biochemistry at Duke University School of Medicine.

As a Principal Investigator in the School of Medicine, his primary role is to keep the lab running (through grant writing and mentoring students), to advocate for his science (by attending and presenting at conferences), and to serve the scientific community (by participating on committees, and specifically, by promoting diversity in STEM at Duke, the School of Medicine, and the American Society of Cellular Biology). Dr. Boyce enjoys his students, and he enjoys mentoring. This can be seen in his lab, where his door is always open for students to stop by to ask questions and to chat.  Finally, he feels very lucky for the freedom offered in his position.  He is able to ask his own questions, and if his project does not work out, or if the results lead to a different question, he is able to follow up.  He believes that his position is as close to a blank check as you can get.

After learning about his path, and what he does, I asked him if he had any advice for people interested in pursuing science. He answered “Stay open minded”, he says that there is a lot to learn, so go to more research seminars (even outside of the realm of expertise!).  Additionally, he recommends to read things that sound interesting, and try to be curiosity driven (because that is what science is about!).  Finally, he explained the importance of forming a network, with people in your lab, your peers, or your professors, because you never know what will happen fifteen years from now.

Dr. Boyce. An awesome mentor and scientist. Original photo was posted on the lab page.

I am thankful to have had the opportunity to sit down and talk to Dr. Boyce about his path, and profession.

On next weeks blog, I will talk about my daily life.

 

 

Week 2- The revelation of my project

Week 2 is complete and as promised in the previous blog post, here are some details for the upcoming project .

To begin, I will give you some background.  In cells, things are always in motion, whether it is RNA leaving the nucleus or proteins transferring from the endoplasmic reticulum (ER) to the Golgi apparatus (Golgi), or vise versa.  For this particular project, we are interested in the forward trafficking, i.e. from the ER to the Golgi.  This forward trafficking pathway is mediated by the Coat protein complex II (COPII).  COPII has five major components:Sar1, Sec13, Sec23, Sec24, and Sec31.  Additionally, posttranslational modification plays an important role in the regulation of the COPII complex. Recently, our lab and others have found that O-linked β-N-acetylglucosamine (O-GlcNAc), a single sugar modification added to serine and threonine residues of intracellular proteins, decorates many COPII components.

Now for the reveal of my project!  I will look at O-GlcNAcylation of Sec24D, a cargo-binding subunit of the COPII complex, because in humans, Sec24D mutations cause a subtype of osteogenesis imperfecta (also known as brittle bone disease), a collagen trafficking disorder. I will do this by answering the question, “What role does Sec24D O-GlcNAcylation play in collagen secretion?”

Here is how I hope to answer the question. I will first create a Sec24D knockout cell line using CRISPR-Cas9 genome editing. Then I will express unglycosylatable Sec24D mutants in the Sec24D knockout cells. Finally, I will determine the role of Sec24D O-GlcNAcylation in collagen secretion by using immunofluorescence microscopy and measuring collagen trafficking to the Golgi.

Here is what I think will happen after this project is complete.  I believe that mutations in Sec24D O-GlcNAc sites will cause collagen secretion to decrease. By doing so, I hope to enhance our understanding of the impacts of O-GlcNAcylation on the function of Sec24D in collagen secretion and allow for future questions on the impacts of O-GlcNAc on other COPII proteins.

Also, check out this paper similar to our project but on Sec23 from our lab.  It is filled with additional background information, and methods that I will be using.

This has been a very exciting two weeks, and I am looking forward to the next six weeks!

On next weeks blog we will have a celebrity interview, so please check it out.

Week 1-Great Anticipation…

During the next eight weeks, I am excited to expand my research experience, that I started last semester, in Dr. Boyce’s lab with my mentor and friend, Brittany.  In the coming weeks, I hope to learn and grow as a person, a member of the Duke community, and a member of the scientific community.

Firstly, I would like to learn to make mistakes.  From my research experience so far, I learned that people make mistakes, and there is a lot of failures. However, I am still hesitant and afraid to make mistakes.  It is my primary goal to get outside of my comfort zone, to try new things, to make mistakes, and to learn from those mistakes.  To add to this goal, I want to learn that it is okay to “fail” and that failing is a part of the process.

Secondly, I would like to learn to be patient.  Science sometimes can be a slow and tedious process.  Sometimes you don’t get what you expect and have to follow up on them.  Sometimes you have just enough downtime so that you can not do anything else other than wait.  The scientific process relies on delayed gratification.  However, I enjoy immediate gratification, therefore I procrastinate ( More on instant gratification and procrastination check out this Ted Talk by Tim Urban), thus, I hope that this opportunity will allow me to practice patience, and appreciate things when they come into fruition.

Finally, I hope to be able to communicate and learn from the community around me.  Through this program, I am fortunate to be able to spend the summer with my peers from BSURF, BioCoRE, and Huang Fellows.  Each individual is taking part in a diverse set of academic research.  As a program, we will have the opportunity to share our research, thus helping with our communication skills, and listen to their research, thus expanding our scientific knowledge.

I am excited to see what this summer will bring and as the summer progresses, I will keep an open mind to learn and grow as much as possible.

That’s it until next week, where I will reveal some exciting details on my summer project.

Western blots are cool.
(Photo was originally posted on the Boyce Lab’s website, check it out!)

 

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