sb7b00440_si_001.pdf (3.46 MB)
Engineering a Functional Small RNA Negative Autoregulation Network with Model-Guided Design
journal contribution
posted on 2018-05-07, 00:00 authored by Chelsea
Y. Hu, Melissa K. Takahashi, Yan Zhang, Julius B. LucksRNA
regulators are powerful components of the synthetic biology
toolbox. Here, we expand the repertoire of synthetic gene networks
built from these regulators by constructing a transcriptional negative
autoregulation (NAR) network out of small RNAs (sRNAs). NAR network
motifs are core motifs of natural genetic networks, and are known
for reducing network response time and steady state signal. Here we
use cell-free transcription–translation (TX-TL) reactions and
a computational model to design and prototype sRNA NAR constructs.
Using parameter sensitivity analysis, we design a simple set of experiments
that allow us to accurately predict NAR function in TX-TL. We transfer
successful network designs into Escherichia coli and
show that our sRNA transcriptional network reduces both network response
time and steady-state gene expression. This work broadens our ability
to construct increasingly sophisticated RNA genetic networks with
predictable function.