PT  - JOURNAL ARTICLE
AU  - David JHF Knapp
AU  - Yale S Michaels
AU  - Max Jamilly
AU  - Quentin RV Ferry
AU  - Hector Barbosa
AU  - Thomas A Milne
AU  - Tudor A Fulga
TI  - Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
AID  - 10.1101/342485
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 342485
4099  - http://biorxiv.org/content/early/2018/06/08/342485.short
4100  - http://biorxiv.org/content/early/2018/06/08/342485.full
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.