This summer, I’m working at the Benaroya Research Institute (BRI), an autoimmune disease research group affiliated with Virginia Mason Hospital in Seattle. I’m in the Lacey-Hulbert Stuart lab, where they mainly study autoimmune gut diseases like IBD and Crohn’s Disease. They also use a fairly new gene identification system to look for genes in human cells that confer resistance to infectious diseases.
I work on the genetic side of things with a postdoc researching resistance to bacterial toxins. Her project started with the identification program, which uses transposon mutagenesis (inserting sections of genes into new parts of the genome to mutate the cell) to randomly generate cell mutations, then moved to infecting the mutated cells with bacterial toxins and sequencing the survivors. One resistance gene that popped up is called LITAF, which appears to be involved in moving material around inside cells. It’s thought that LITAF may promote toxin resistance by trapping the toxins in little bubbles of cell membrane called endosomes and then destroying these toxins or expelling them from the cell before they can do much damage.
This is where I joined the project. Right now, we’re trying to refute some possibilities other researchers have raised that LITAF may, instead of moving toxins quickly out of the cell, prevent toxins from getting past the cell membrane in the first place. The experiment I’m running today will hopefully disprove that possibility. I’m doing something called a kinetic stress test—adding a toxin to cells that usually kills them after a while and measuring markers of cell stress that the cells spit out as they start to die. I got here at 8:30 am to set up the experiment and do some housekeeping: warming up the liquid we grow cells in, checking cell samples under the microscope, moving some bacteria we’re growing from tiny test tubes to big bottles where they can happily multiply, putting virus on cells we want to study later, and generally making sure everything is in order. We want to know how stressed the cells become with different exposure times to toxin, so I added toxin to the cells at five different times today, from 10 am to 2 pm at shortening intervals (I have a timer to keep me on track). This way we end up with samples of cells that have been sitting in toxin for 4 hours, 2 hours, 1 hour, 30 minutes, and 15 minutes.
In between adding toxin, I took a look at some fluorescent-stained cells in a machine that costs way more than my net worth and tried really hard not to mess anything up, then went out to lunch (a sushi place a few blocks from the lab) with a group of postdocs and a guest speaker from Carnegie Mellon who came to give a lecture at a cell imaging conference down the street on new genetic tagging techniques using synthetic dyes. We talked about his research, but I also got to hear about maintaining lab cohesion, everyday research troubles, how different institutes run graduate school programs, followed by a lengthy discussion on the 80s and Ferris Bueller’s Day Off.
After that break, I kept to my timer and added toxin to my cells according to schedule while writing up the day’s work in my lab notebook and putting next week’s immunology lectures and lab meetings into my schedule. I heard a little bit about another postdoc’s Ebola project, then hurried over to the tissue culture room (where we work with human cells, blood, and infectious material) at 2 pm to collect the cells from the kinetic stress test and freeze them for imaging later. I spent the rest of the afternoon finishing up a PCR test on some samples from an old experiment, wrote out my experiment plan for the next day, then walked back down First Hill, past the Central Library, and down towards the waterfront where I caught a bus to take me home.