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

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 decrease...

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Autores principales: Sarkar, Mrinal K., Uppala, Ranjitha, Zeng, Chang, Billi, Allison C., Tsoi, Lam C., Kidder, Austin, Xing, Xianying, Perez White, Bethany E., Shao, Shuai, Plazyo, Olesya, Sirobhushanam, Sirisha, Xing, Enze, Jiang, Yanyun, Gallagher, Katherine A., Voorhees, John J., Kahlenberg, J. Michelle, Gudjonsson, Johann E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Clinical Investigation 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10145927/
https://www.ncbi.nlm.nih.gov/pubmed/36928117
http://dx.doi.org/10.1172/JCI159393
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author Sarkar, Mrinal K.
Uppala, Ranjitha
Zeng, Chang
Billi, Allison C.
Tsoi, Lam C.
Kidder, Austin
Xing, Xianying
Perez White, Bethany E.
Shao, Shuai
Plazyo, Olesya
Sirobhushanam, Sirisha
Xing, Enze
Jiang, Yanyun
Gallagher, Katherine A.
Voorhees, John J.
Kahlenberg, J. Michelle
Gudjonsson, Johann E.
author_facet Sarkar, Mrinal K.
Uppala, Ranjitha
Zeng, Chang
Billi, Allison C.
Tsoi, Lam C.
Kidder, Austin
Xing, Xianying
Perez White, Bethany E.
Shao, Shuai
Plazyo, Olesya
Sirobhushanam, Sirisha
Xing, Enze
Jiang, Yanyun
Gallagher, Katherine A.
Voorhees, John J.
Kahlenberg, J. Michelle
Gudjonsson, Johann E.
author_sort Sarkar, Mrinal K.
collection PubMed
description 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 gene 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 for 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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spelling pubmed-101459272023-05-01 Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction Sarkar, Mrinal K. Uppala, Ranjitha Zeng, Chang Billi, Allison C. Tsoi, Lam C. Kidder, Austin Xing, Xianying Perez White, Bethany E. Shao, Shuai Plazyo, Olesya Sirobhushanam, Sirisha Xing, Enze Jiang, Yanyun Gallagher, Katherine A. Voorhees, John J. Kahlenberg, J. Michelle Gudjonsson, Johann E. J Clin Invest Research Article 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 gene 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 for 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. American Society for Clinical Investigation 2023-05-01 /pmc/articles/PMC10145927/ /pubmed/36928117 http://dx.doi.org/10.1172/JCI159393 Text en © 2023 Sarkar et al. https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Sarkar, Mrinal K.
Uppala, Ranjitha
Zeng, Chang
Billi, Allison C.
Tsoi, Lam C.
Kidder, Austin
Xing, Xianying
Perez White, Bethany E.
Shao, Shuai
Plazyo, Olesya
Sirobhushanam, Sirisha
Xing, Enze
Jiang, Yanyun
Gallagher, Katherine A.
Voorhees, John J.
Kahlenberg, J. Michelle
Gudjonsson, Johann E.
Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title_full Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title_fullStr Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title_full_unstemmed Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title_short Keratinocytes sense and eliminate CRISPR DNA through STING/IFN-κ activation and APOBEC3G induction
title_sort keratinocytes sense and eliminate crispr dna through sting/ifn-κ activation and apobec3g induction
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10145927/
https://www.ncbi.nlm.nih.gov/pubmed/36928117
http://dx.doi.org/10.1172/JCI159393
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