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Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization

Author

Listed:
  • Serif Senturk

    (Cold Spring Harbor Laboratory)

  • Nitin H. Shirole

    (Cold Spring Harbor Laboratory
    Graduate Program in Genetics, Stony Brook University)

  • Dawid G. Nowak

    (Cold Spring Harbor Laboratory)

  • Vincenzo Corbo

    (Cold Spring Harbor Laboratory)

  • Debjani Pal

    (Cold Spring Harbor Laboratory
    Graduate Program in Molecular and Cellular Biology, Stony Brook University)

  • Alexander Vaughan

    (Cold Spring Harbor Laboratory)

  • David A. Tuveson

    (Cold Spring Harbor Laboratory)

  • Lloyd C. Trotman

    (Cold Spring Harbor Laboratory)

  • Justin B. Kinney

    (Cold Spring Harbor Laboratory)

  • Raffaella Sordella

    (Cold Spring Harbor Laboratory)

Abstract

The CRISPR/Cas9 system is a powerful tool for studying gene function. Here, we describe a method that allows temporal control of CRISPR/Cas9 activity based on conditional Cas9 destabilization. We demonstrate that fusing an FKBP12-derived destabilizing domain to Cas9 (DD-Cas9) enables conditional Cas9 expression and temporal control of gene editing in the presence of an FKBP12 synthetic ligand. This system can be easily adapted to co-express, from the same promoter, DD-Cas9 with any other gene of interest without co-modulation of the latter. In particular, when co-expressed with inducible Cre-ERT2, our system enables parallel, independent manipulation of alleles targeted by Cas9 and traditional recombinase with single-cell specificity. We anticipate this platform will be used for the systematic characterization and identification of essential genes, as well as the investigation of the interactions between functional genes.

Suggested Citation

  • Serif Senturk & Nitin H. Shirole & Dawid G. Nowak & Vincenzo Corbo & Debjani Pal & Alexander Vaughan & David A. Tuveson & Lloyd C. Trotman & Justin B. Kinney & Raffaella Sordella, 2017. "Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization," Nature Communications, Nature, vol. 8(1), pages 1-10, April.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms14370
    DOI: 10.1038/ncomms14370
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    Cited by:

    1. Xing Cheng & Jing An & Jitong Lou & Qisheng Gu & Weimin Ding & Gaith Nabil Droby & Yilin Wang & Chenghao Wang & Yanzhe Gao & Jay Ramanlal Anand & Abigail Shelton & Andrew Benson Satterlee & Breanna Ma, 2024. "Trans-lesion synthesis and mismatch repair pathway crosstalk defines chemoresistance and hypermutation mechanisms in glioblastoma," Nature Communications, Nature, vol. 15(1), pages 1-20, December.

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