@article {Port046417, author = {Fillip Port and Simon L Bullock}, title = {Expansion of the CRISPR toolbox in an animal with tRNA-flanked Cas9 and Cpf1 gRNAs}, elocation-id = {046417}, year = {2016}, doi = {10.1101/046417}, publisher = {Cold Spring Harbor Laboratory}, abstract = {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 {\textdagger}RNAs, permits highly efficient multiplexing of Cas9-based mutagenesis. We also demonstrate that the {\textdagger}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.}, URL = {https://www.biorxiv.org/content/early/2016/03/31/046417.1}, eprint = {https://www.biorxiv.org/content/early/2016/03/31/046417.1.full.pdf}, journal = {bioRxiv} }