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Multiplexed CRISPR technologies for gene editing and transcriptional regulation

Author

Listed:
  • Nicholas S. McCarty

    (California Institute of Technology)

  • Alicia E. Graham

    (Imperial College London)

  • Lucie Studená

    (Imperial College London)

  • Rodrigo Ledesma-Amaro

    (Imperial College London)

Abstract

Multiplexed CRISPR technologies, in which numerous gRNAs or Cas enzymes are expressed at once, have facilitated powerful biological engineering applications, vastly enhancing the scope and efficiencies of genetic editing and transcriptional regulation. In this review, we discuss multiplexed CRISPR technologies and describe methods for the assembly, expression and processing of synthetic guide RNA arrays in vivo. Applications that benefit from multiplexed CRISPR technologies, including cellular recorders, genetic circuits, biosensors, combinatorial genetic perturbations, large-scale genome engineering and the rewiring of metabolic pathways, are highlighted. We also offer a glimpse of emerging challenges and emphasize experimental considerations for future studies.

Suggested Citation

  • Nicholas S. McCarty & Alicia E. Graham & Lucie Studená & Rodrigo Ledesma-Amaro, 2020. "Multiplexed CRISPR technologies for gene editing and transcriptional regulation," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15053-x
    DOI: 10.1038/s41467-020-15053-x
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    Cited by:

    1. Yuting Chen & Eriona Hysolli & Anlu Chen & Stephen Casper & Songlei Liu & Kevin Yang & Chenli Liu & George Church, 2022. "Multiplex base editing to convert TAG into TAA codons in the human genome," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    2. Sally Wang & Chen-Yu Chen & John R. Rzasa & Chen-Yu Tsao & Jinyang Li & Eric VanArsdale & Eunkyoung Kim & Fauziah Rahma Zakaria & Gregory F. Payne & William E. Bentley, 2023. "Redox-enabled electronic interrogation and feedback control of hierarchical and networked biological systems," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    3. Marco Thürkauf & Shuo Lin & Filippo Oliveri & Dirk Grimm & Randall J. Platt & Markus A. Rüegg, 2023. "Fast, multiplexable and efficient somatic gene deletions in adult mouse skeletal muscle fibers using AAV-CRISPR/Cas9," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    4. Jianli Tao & Daniel E. Bauer & Roberto Chiarle, 2023. "Assessing and advancing the safety of CRISPR-Cas tools: from DNA to RNA editing," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    5. Guanhua Xun & Zhixin Zhu & Nilmani Singh & Jingxia Lu & Piyush K. Jain & Huimin Zhao, 2024. "Harnessing noncanonical crRNA for highly efficient genome editing," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    6. Dixit, Yatika & Yadav, Preeti & Sharma, Arun Kumar & Pandey, Poornima & Kuila, Arindam, 2023. "Multiplex genome editing to construct cellulase engineered Saccharomyces cerevisiae for ethanol production from cellulosic biomass," Renewable and Sustainable Energy Reviews, Elsevier, vol. 187(C).
    7. Jessica A. Kretzmann & Anna Liedl & Alba Monferrer & Volodymyr Mykhailiuk & Samuel Beerkens & Hendrik Dietz, 2023. "Gene-encoding DNA origami for mammalian cell expression," Nature Communications, Nature, vol. 14(1), pages 1-10, December.
    8. Qichen Yuan & Xue Gao, 2022. "Multiplex base- and prime-editing with drive-and-process CRISPR arrays," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    9. William M. Shaw & Lucie Studená & Kyler Roy & Piotr Hapeta & Nicholas S. McCarty & Alicia E. Graham & Tom Ellis & Rodrigo Ledesma-Amaro, 2022. "Inducible expression of large gRNA arrays for multiplexed CRISPRai applications," Nature Communications, Nature, vol. 13(1), pages 1-10, December.

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