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.