Plasmid Lineages

Recombinant DNA work is integral to what we’re doing here, so I’ve become extremely organized with keeping track of the constructs we are building. This includes having a record of how sequences from two constructs were stitched together to create a new construct. Here’s a network map showing how one or more different plasmid sequences were combined to create each new construct.

[The series of letters and numbers prefixed with G (for Gibson) are unique identifiers I started giving new constructs when it became clear partway through my postdoc that I was going to need a better way of tracking everything I was building. Those prefixed with A are constructs obtained through addgene. Those prefixed with R are important constructs I had built before this tracking system, where I had to start giving them identifiers retroactively.]

HEK293Ts with melanin

I think synthetic biology is really cool, and I like playing around with recombinant DNA elements so I can see how well they work in my own hands. If they work OK, then I just let that knowledge stew in the back of my brain until I can eventually figure out a use for it. Reading this paper by Martin Fussenegger made me realize just how easy it is to make cultured cells express melanin. Here was my first foray in creating melanin in HEK cells by overexpressing tyrosinase/

Cells pelleted in the tubes on the left are expressing tyrosinase. The cells pelleted in the tubes on the right are not.

Doesn’t quite work well enough to use as a general reporter (it’s really hard to tell in a cell monolayer, and only becomes noticeable as colonies of cells or in a pellet, like above), but still kind of fun to see. Let’s see if I find an eventual use for this in some future work.