🚨New paper alert!🚨 Cells integrate signals into decision-making. But how do levels of signaling influence cell-fate transitions? Is more always better? Or is there an “optimal” level for cell-fate programming? If we could tune signaling, could optimize production of neurons? Read on! (1/n)🧵 To ask this question, Brittany started probing the the MAPK pathway. The MAPK pathway is a central regulator of cellular decision-making. The levels and dynamics of signaling direct cells down different fate. Based on previous work, we knew that adding a MAPK mutant (HRAS-G12V) increases direct conversion of skin cells to motor neuron when combined motor neuron TFs (NIL) and a p53 mutant. (2/n)

So you want to change transgene expression: just change your promoter, right? Changing the promoter increases RNA and thus protein levels. What more could be happening?

[1/n] Well, promoters don’t just set RNA levels; they uniquely transform how RNAs are transmitted into protein levels. 🧵 [2/n] Buckle up for this delightful journey through the Central Dogma led by @emmapeterman_ and now published at NAR on how promoters change expression...but not in the way we were expecting.

Article: doi.org/10.1093/nar/gk…

So you want to engineer your hiPSCs, but targeting DNA payloads requires multiple slow, inefficient steps for each construct. What if we could accomplish multi-site integration seamlessly?
🚨Introducing STRAIGHT-IN Dual! 🧵 (1/n)
Link: biorxiv.org/content/10.110… STRAIGHT-IN Dual enables simultaneous, allele-specific, single-copy integration of two DNA payloads with 100% efficiency within one week. Engineering the landing pad (LP) with a promoter trap supports 100% efficiency through selection (e.g. burn-the-hay strategy). (2/n)

Loss of gene expression can compromise cellular fitness. In the case of such haploinsufficiency, replacement of the gene supplementation could rescue cell health. But what if you need a very precise dosage? Try ComMAND! 🧵 (1/n)
Link: doi.org/10.1101/2024.0… Compact microRNA-Mediated Attenuator of Noise and Dosage (ComMAND), is a single-transcript microRNA incoherent feedforward loop, which means it constrains transgene expression through production of a transcript-encoded, self-targeting microRNA.(2/n)

Levels of gene expression matter! So how can you control a transgene precisely? Is there a tool that can ensure uniform control & the ability to adjust the setpoint of expression from a single promoter? Introducing DIAL! 🧵 (1/n)
Link: doi.org/10.1101/2024.0… What is DIAL? DIAL is a framework (and a toolkit) for designing synthetic promoters that generate heritable, unimodal setpoints of gene expression from a single promoter...(2/n)

How do individual transcription factors (TF) influence reprogramming? Isn’t it just the cells with the highest levels of TF expression that reprogram? So just blast them & it should work, right? 🤨Buckle up for our lab’s preprint & tour de force led by Nat Wang @nbwang22 ! (1/n)

https://twitter.com/biorxiv_cellbio/status/1729206136476209330

On its own, high expression of transcription isn’t a great predictor of reprogramming outcomes. So what else do cells need? (2/n)

How does transcription of proximal genes influence the expression of their neighbors? How do we engineer compact circuit designs for specific profiles of expression that enable robust cellular engineering? Today @CellReports we provide some answers (1 /n) tinyurl.com/24vtfbyw We integrated DNA biophysics with classic #synbio and stochastic simulations to explore how to optimize the expression levels, variance, and dynamic response of transgenic systems. We found that supercoiling-mediated feedback significantly influences these behaviors! (2/n)

Research experience is so formative for undergrads, but right now in-person training isn't an option for most. During the shutdown, we switched to emergency computational modeling (surprisingly successful!) This semester we're taking a more proactive training approach...(1/n) ...and experimenting with a series of didactic trainings on reading lit, reviewing lit, writing proposals, and building graphics and figures. And then having the students deliver these items. While these are essential skills, I don't recall being taught as a UG to do this...(2/n)