Competitive dCas9 binding as a mechanism for transcriptional control
Name
msb.202110512.pdf
Description
Published version
Size
1015.91 KB
Format
Adobe PDF
Checksum (MD5)
dfc15fe5152a38d58ebe755cdf5e4d69
Author(s)
Anderson, Daniel A
Voigt, Christopher A
Date Issued
November 2021
Journal
Molecular Systems Biology
Publisher
EMBO
Citation
Anderson, Daniel A and Voigt, Christopher A. 2021. "Competitive dCas9 binding as a mechanism for transcriptional control." Molecular Systems Biology, 17 (11).
Version
Final published version
Abstract
Catalytically dead Cas9 (dCas9) is a programmable transcription factor that can be targeted to promoters through the design of small guide RNAs (sgRNAs), where it can function as an activator or repressor. Natural promoters use overlapping binding sites as a mechanism for signal integration, where the binding of one can block, displace, or augment the activity of the other. Here, we implemented this strategy in Escherichia coli using pairs of sgRNAs designed to repress and then derepress transcription through competitive binding. When designed to target a promoter, this led to 27-fold repression and complete derepression. This system was also capable of ratiometric input comparison over two orders of magnitude. Additionally, we used this mechanism for promoter sequence-independent control by adopting it for elongation control, achieving 8-fold repression and 4-fold derepression. This work demonstrates a new genetic control mechanism that could be used to build analog circuit or implement cis-regulatory logic on CRISPRi-targeted native genes.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.15252/msb.202110512
