Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs.

ABSTRACT: We present tRNA-based vectors for producing multiple clustered regularly interspaced short palindromic repeats (CRISPR) single guide RNAs (sgRNAs) from a single RNA polymerase II or III transcript in Drosophila. The system, which is based on liberation of sgRNAs by processing flanking tRNAs, permits highly efficient multiplexing of Cas9-based mutagenesis. We also demonstrate that the tRNA-sgRNA system markedly increases the efficacy of conditional gene disruption by Cas9 and can promote editing by the recently discovered RNA-guided endonuclease Cpf1.

SUBMITTER: Port F 

PROVIDER: S-EPMC5215823 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature