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Nat Methods. 2016 Oct;13(10):852-4. doi: 10.1038/nmeth.3972. Epub 2016 Sep 5.

Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs.

Author information

1
Cell Biology Division, MRC Laboratory of Molecular Biology, Cambridge, UK.

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.

PMID:
27595403
PMCID:
PMC5215823
DOI:
10.1038/nmeth.3972
[Indexed for MEDLINE]
Free PMC Article

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