RT Journal Article
SR Electronic
T1 Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 342485
DO 10.1101/342485
A1 Knapp, David JHF
A1 Michaels, Yale S
A1 Jamilly, Max
A1 Ferry, Quentin RV
A1 Barbosa, Hector
A1 Milne, Thomas A
A1 Fulga, Tudor A
YR 2018
UL http://biorxiv.org/content/early/2018/06/08/342485.abstract
AB Spatial/temporal control of Cas9 guide RNA expression could considerably expand the utility of CRISPR-based technologies. Current approaches based on tRNA processing offer a promising strategy but suffer from high background. Here we developed a variant screening platform to identify differential sequence determinants of human tRNA promoter and processing activities. Rational design based on the ensuing principles allowed us to engineer an improved tRNA scaffold that enabled highly specific guide RNA production from a Pol-II promoter.