Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs

@article{Port2016AugmentingCA,
  title={Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs},
  author={Fillip Port and Simon L Bullock},
  journal={Nature Methods},
  year={2016},
  volume={13},
  pages={852-854}
}
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… CONTINUE READING
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