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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

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Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs.

Port Fillip F Bullock Simon L SL

Nature methods 20160905 10

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 ...[more]

PMID: 27595403

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