It's a crazy time to be a scientist, but I feel fortunate and excited to be able to share the first bit of work from my postdoc... introducing PRANCE!

"A high-throughput platform for feedback-controlled directed evolution"

Here's the rundown... 1/n

https://www.biorxiv.org/content/10.1101/2020.04.01.021022v1

I am so proud to have been a part of this project in the @kesvelt Sculpting Evolution Lab with an incredible team: 1st author/rockstar @erika_alden_d and @gretton_dana @bscwang

First, PRANCE = Phage-and-robot-assisted near-continuous evolution ….. 2/n

So... we developed a method to do PACE in high-throughput with real-time luminescence reporter monitoring using some fancy robotics and an EPIC custom python package dev'd by @gretton_dana (stay tuned for our python paper in the coming weeks! )…. 3/n

After validating the platform every which way, we first ran ONE HUNDRED simultaneous T7RNAP evolutions.



*AGASP* at how normally distributed evolution is?!?? We were too! …. 4/n

The super cool thing about PRANCE is that it miniaturizes PACE 100-fold, so we can actually run evolutions with expensive small molecules, like custom non-canonical amino acids! With replicates! And controls!…. 5/n

Ever wanted to evolve a quadruplet-decoding tRNA for all 20 amino acids... simultaneously? Now you can! And we did…. 6/n

It turns out that one of the hardest things about evolution is that proteins all have different starting activities i.e. phage can easily wash out, or the evolution could be too strong being with.

So, how do you know what to do or what experiment to run?…..well.... 7/n

With real-time monitoring, we developed feedback-control that automatically bumps up the bacteria stringency when we sense phage, but not before.

Now you can run dozens of evolutions simultaneously, even with different bacteria and different starting activities!!… 8/n

So basically with PRANCE, we were able to run hundreds of different evolutions (in parallel) with 100-fold less reagents in just a few experiments. This work summarizes a massive effort and I could not be prouder of the team…. 9/n

We’d love any feedback you have, support, or ideas. It's crazy to think that just a few months ago, we were still fighting clogged bleach lines and robot collisions. Now it feels like the world is our oyster and I can’t wait for what’s next…. 10/n

Special thanks to our brilliant and supportive mentor @kesvelt and congrats to the whole team, but especially @erika_alden_d on her FIRST first author paper from grad school. Keep an eye on this one, she’s going places…. fin.

TBH to one of our 96-PRANCE experiments https://twitter.com/chorye/status/1230291803540090880?s=20