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Cpf1 proteins excise CRISPR RNAs from mRNA transcripts in mammalian cells

Nature Chemical Biology volume 13, pages 839–841 (2017)Cite this article

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Abstract

Cpf1 is a CRISPR effector protein that has greater specificity than Streptococcus pyogenes Cas9 (SpCas9) in genome-editing applications. Here we show that Lachnospiraceae bacterium (Lb) and Acidaminococus sp. (As) Cpf1 orthologs have RNase activities that can excise multiple CRISPR RNAs (crRNAs) from a single RNA polymerase II–driven RNA transcript expressed in mammalian cells. This property simplifies modification of multiple genomic targets and can be used to increase the efficiency of Cpf1-mediated editing.

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Figure 1: LbCpf1 and AsCpf1 have RNase activities in mammalian cells.
Figure 2: crRNA excised from a Pol II–expressed RNA transcript can efficiently edit a mammalian genome.
Figure 3: Cpf1 efficiently excises and uses tandemly arrayed crRNAs expressed from a Pol II promoter.

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References

  1. Zetsche, B. et al. Cell 163, 759–771 (2015).

    Article  CAS  Google Scholar 

  2. Kim, D. et al. Nat. Biotechnol. 34, 863–868 (2016).

    Article  CAS  Google Scholar 

  3. Kleinstiver, B.P. et al. Nat. Biotechnol. 34, 869–874 (2016).

    Article  CAS  Google Scholar 

  4. Mohanraju, P. et al. Science 353, aad5147 (2016).

    Article  Google Scholar 

  5. Fonfara, I., Richter, H., Bratovič, M., Le Rhun, A. & Charpentier, E. Nature 532, 517–521 (2016).

    Article  CAS  Google Scholar 

  6. Dong, D. et al. Nature 532, 522–526 (2016).

    Article  CAS  Google Scholar 

  7. Yamano, T. et al. Cell 165, 949–962 (2016).

    Article  CAS  Google Scholar 

  8. Certo, M.T. et al. Nat. Methods 8, 671–676 (2011).

    Article  CAS  Google Scholar 

  9. Orioli, A. et al. Nucleic Acids Res. 39, 5499–5512 (2011).

    Article  CAS  Google Scholar 

  10. Shcherbakova, D.M. & Verkhusha, V.V. Nat. Methods 10, 751–754 (2013).

    Article  CAS  Google Scholar 

  11. Pédelacq, J.D., Cabantous, S., Tran, T., Terwilliger, T.C. & Waldo, G.S. Nat. Biotechnol. 24, 79–88 (2006).

    Article  Google Scholar 

  12. Zetsche, B. et al. Nat. Biotechnol. 35, 31–34 (2017).

    Article  CAS  Google Scholar 

  13. Samulski, R.J. & Muzyczka, N. Annu. Rev. Virol. 1, 427–451 (2014).

    Article  Google Scholar 

  14. Guschin, D.Y. et al. Methods Mol. Biol. 649, 247–256 (2010).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work is supported by NIH R37 AI091476 and P01 AI100263 (M.F.).

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Authors and Affiliations

  1. Department of Immunology and Microbiology, The Scripps Research Institute, Jupiter, Florida, USA

    Guocai Zhong, Haimin Wang, Yujun Li, Mai H Tran & Michael Farzan

  2. Department of Microbiology and Wu Lien-Teh Institute, Harbin Medical University, Harbin, China

    Yujun Li

Authors
  1. Guocai Zhong
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  2. Haimin Wang
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  3. Yujun Li
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  4. Mai H Tran
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  5. Michael Farzan
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Contributions

G.Z. conceived this study and developed it with assistance from M.F.; G.Z., H.W., Y.L., and M.H.T. performed all experiments; M.F. and G.Z. wrote the manuscript.

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Correspondence to Guocai Zhong.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Tables 1–4 and Supplementary Figures 1–6 (PDF 8474 kb)

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Zhong, G., Wang, H., Li, Y. et al. Cpf1 proteins excise CRISPR RNAs from mRNA transcripts in mammalian cells. Nat Chem Biol 13, 839–841 (2017). https://doi.org/10.1038/nchembio.2410

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