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Photoactivatable Cre recombinase 3.0 for in vivo mouse applications

Author

Listed:
  • Kumi Morikawa

    (Columbia University
    Columbia University
    National Institute of Advanced Industrial Science and Technology)

  • Kazuhiro Furuhashi

    (Columbia University
    Columbia University)

  • Carmen Sena-Tomas

    (Columbia University
    Columbia University)

  • Alvaro L. Garcia-Garcia

    (Columbia University)

  • Ramsey Bekdash

    (Columbia University
    Columbia University
    Columbia University)

  • Alison D. Klein

    (Columbia University
    Columbia University)

  • Nicholas Gallerani

    (Columbia University
    Columbia University)

  • Hannah E. Yamamoto

    (Columbia University
    Columbia University
    Barnard College)

  • Seon-Hye E. Park

    (Columbia University
    Columbia University
    University of Texas Southwestern Medical Center)

  • Grant S. Collins

    (Louisiana State University Health Sciences Center)

  • Fuun Kawano

    (Columbia University
    Columbia University
    The University of Tokyo)

  • Moritoshi Sato

    (The University of Tokyo
    Japan Science and Technology Agency)

  • Chyuan-Sheng Lin

    (Columbia University
    Columbia University)

  • Kimara L. Targoff

    (Columbia University
    Columbia University)

  • Edmund Au

    (Columbia University
    Columbia University
    Columbia University Irving Medical Center)

  • Michael C. Salling

    (Louisiana State University Health Sciences Center
    Columbia University)

  • Masayuki Yazawa

    (Columbia University
    Columbia University
    Columbia University
    Japan Science and Technology Agency)

Abstract

Optogenetic genome engineering tools enable spatiotemporal control of gene expression and provide new insight into biological function. Here, we report the new version of genetically encoded photoactivatable (PA) Cre recombinase, PA-Cre 3.0. To improve PA-Cre technology, we compare light-dimerization tools and optimize for mammalian expression using a CAG promoter, Magnets, and 2A self-cleaving peptide. To prevent background recombination caused by the high sequence similarity in the dimerization domains, we modify the codons for mouse gene targeting and viral production. Overall, these modifications significantly reduce dark leak activity and improve blue-light induction developing our new version, PA-Cre 3.0. As a resource, we have generated and validated AAV-PA-Cre 3.0 as well as two mouse lines that can conditionally express PA-Cre 3.0. Together these new tools will facilitate further biological and biomedical research.

Suggested Citation

  • Kumi Morikawa & Kazuhiro Furuhashi & Carmen Sena-Tomas & Alvaro L. Garcia-Garcia & Ramsey Bekdash & Alison D. Klein & Nicholas Gallerani & Hannah E. Yamamoto & Seon-Hye E. Park & Grant S. Collins & Fu, 2020. "Photoactivatable Cre recombinase 3.0 for in vivo mouse applications," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-16030-0
    DOI: 10.1038/s41467-020-16030-0
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    Cited by:

    1. Charlotte Cautereels & Jolien Smets & Jonas De Saeger & Lloyd Cool & Yanmei Zhu & Anna Zimmermann & Jan Steensels & Anton Gorkovskiy & Thomas B. Jacobs & Kevin J. Verstrepen, 2024. "Orthogonal LoxPsym sites allow multiplexed site-specific recombination in prokaryotic and eukaryotic hosts," Nature Communications, Nature, vol. 15(1), pages 1-15, December.
    2. Riccardo De Santis & Fred Etoc & Edwin A. Rosado-Olivieri & Ali H. Brivanlou, 2021. "Self-organization of human dorsal-ventral forebrain structures by light induced SHH," Nature Communications, Nature, vol. 12(1), pages 1-11, December.
    3. Miguel Vizoso & Colin E. J. Pritchard & Lorenzo Bombardelli & Bram van den Broek & Paul Krimpenfort & Roderick L. Beijersbergen & Kees Jalink & Jacco van Rheenen, 2022. "A doxycycline- and light-inducible Cre recombinase mouse model for optogenetic genome editing," Nature Communications, Nature, vol. 13(1), pages 1-15, December.

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