RT Journal Article
SR Electronic
T1 Modeling predicts that CRISPR-based activators, unlike CRISPR-based repressors, scale well with increasing gRNA competition and dCas9 bottlenecking
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 719278
DO 10.1101/719278
A1 Clamons, Samuel
A1 Murray, Richard
YR 2019
UL http://biorxiv.org/content/early/2019/08/13/719278.abstract
AB Synthetic transcriptional networks built from CRISPR-based repressors (CRISPRi) rely on shared use of a core dCas9 protein. In E. coli, CRISPRi cannot support more than about a dozen simultaneous gRNAs before the fold repression of any individual gRNA drops below 10x. We show with a simple model based on previous characterization of competition in CRISPRi that activation by CRISPR-based activators (CRISPRa) is much less sensitive to dCas9 bottle-necking than CRISPRi. We predict that E. coli should be able to support dozens to hundreds of CRISPRa gRNAs at >10-fold activation.