Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs.
basic_science · Level V
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- Record sourced from PubMed, PMID 27595403.
- Also identified by DOI 10.1038/nmeth.3972 and PMC identifier 5215823.
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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.
Medical subject headings
- CRISPR-Associated Proteins
- Clustered Regularly Interspaced Short Palindromic Repeats
- Drosophila
- RNA, Guide, CRISPR-Cas Systems
- RNA, Transfer