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Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs
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Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs
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Journal Article
Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs
2016
Overview
Expressing guide RNAs from a tRNA scaffold enhances mutagenesis by both Cas9 and Cpf1 and allows conditional CRISPR applications
in vivo
.
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.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Animals
/ Animals, Genetically Modified - genetics
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Clustered Regularly Interspaced Short Palindromic Repeats - genetics
/ CRISPR-Associated Proteins - genetics
/ CRISPR-Cas Systems - genetics
/ Insects
/ Plasmids
/ RNA Polymerase II - genetics
/ RNA Polymerase III - genetics
