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ResearchIn-Press PreviewCell biologyDermatology Open Access | 10.1172/JCI159393

Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction

Mrinal K. Sarkar,1 Ranjitha Uppala,1 Chang Zeng,1 Allison C. Billi,1 Lam C. Tsoi,2 Austin Kidder,1 Xianying Xing,1 Bethany E. Perez White,3 Shuai Shao,4 Olesya Plazyo,1 Sirisha Sirobhushanam,5 Enze Xing,1 Yanyun Jiang,6 Katherine A. Gallagher,7 John J. Voorhees,1 J. Michelle Kahlenberg,5 and Johann E. Gudjonsson2

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Sarkar, M. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Uppala, R. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Zeng, C. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Billi, A. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Tsoi, L. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Kidder, A. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Xing, X. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Perez White, B. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Shao, S. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Plazyo, O. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Sirobhushanam, S. in: JCI | PubMed | Google Scholar

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Xing, E. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Jiang, Y. in: JCI | PubMed | Google Scholar

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Gallagher, K. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Voorhees, J. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Kahlenberg, J. in: JCI | PubMed | Google Scholar |

1Department of Dermatology, University of Michigan, Ann Arbor, United States of America

2University of Michigan, Ann Arbor, United States of America

3Department of Dermatology, Northwestern University, Chicago, United States of America

4Department of Dermatology, Xijing Hospital, Fourth Military Medical Univers, Xi’an, China

5Department of Internal Medicine, Division of Rheumatology, University of Michigan, Ann Arbor, United States of America

6Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China

7Department of Vascular Surgery, University of Michigan, Ann Arbor, United States of America

Find articles by Gudjonsson, J. in: JCI | PubMed | Google Scholar |

Published March 16, 2023 - More info

J Clin Invest. https://doi.org/10.1172/JCI159393.
Copyright © 2023, Sarkar et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published March 16, 2023 - Version history
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Abstract

CRISPR-Cas9 has been proposed as a treatment for genetically inherited skin disorders. Here we report that CRISPR transfection activates STING-dependent antiviral responses in keratinocytes, resulting in heightened endogenous interferon (IFN) responses through induction of IFN-κ leading to decreased plasmid stability secondary to induction of the cytidine deaminase APOBEC3G. Notably, CRISPR-generated KO keratinocytes had permanent suppression of IFN-κ and IFN-stimulated gene (ISG) expression, secondary to hypermethylation of the IFNK promoter region by the DNA methyltransferase DNMT3B. JAK inhibition via baricitinib prior to CRISPR transfection increased transfection efficiency, prevented IFNK promoter hypermethylation, and restored normal IFN-κ activity and ISG responses. This work shows that CRISPR-mediated gene correction alters antiviral responses in keratinocytes, has implications for future gene therapies of inherited skin diseases using CRISPR technology, and suggests pharmacologic JAK inhibition as a tool for facilitating and attenuating inadvertent selection effects in CRISPR-Cas9 therapeutic approaches.

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