%0 Journal Article %A Fillip Port %A Simon L Bullock %T Expansion of the CRISPR toolbox in an animal with tRNA-flanked Cas9 and Cpf1 gRNAs %D 2016 %R 10.1101/046417 %J bioRxiv %P 046417 %X We present vectors for producing multiple CRISPR gRNAs from a single RNA polymerase II or III transcript in Drosophila. The system, which is based on liberation of gRNAs by processing of flanking t †RNAs, permits highly efficient multiplexing of Cas9-based mutagenesis. We also demonstrate that the †RNA-gRNA system markedly increases the efficacy of conditional gene disruption by Cas9 and can promote editing by the recently discovered RNA-guided endonuclease Cpf1. %U https://www.biorxiv.org/content/biorxiv/early/2016/03/31/046417.1.full.pdf