Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model

Autoimmunity-induced pancreatic beta cell failure is the main characteristic of type 1 diabetes (T1D). Here, we describe a protocol for genome-scale in vivo CRISPR-Cas9 screening for use in a mouse model of T1D. Using a non-obese-diabetic-derived mouse beta cell line, NIT-1, and a genome-wide CRISPR...

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Autores principales: Li, Jian, Lee, Yu-Chi, Iessi, Isabela L., Wu, Chialing, Yi, Peng, Cai, Erica P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025263/
https://www.ncbi.nlm.nih.gov/pubmed/36917606
http://dx.doi.org/10.1016/j.xpro.2023.102155
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author Li, Jian
Lee, Yu-Chi
Iessi, Isabela L.
Wu, Chialing
Yi, Peng
Cai, Erica P.
author_facet Li, Jian
Lee, Yu-Chi
Iessi, Isabela L.
Wu, Chialing
Yi, Peng
Cai, Erica P.
author_sort Li, Jian
collection PubMed
description Autoimmunity-induced pancreatic beta cell failure is the main characteristic of type 1 diabetes (T1D). Here, we describe a protocol for genome-scale in vivo CRISPR-Cas9 screening for use in a mouse model of T1D. Using a non-obese-diabetic-derived mouse beta cell line, NIT-1, and a genome-wide CRISPR-Cas9 knockout library (GeCKO-v2), we describe how to identify genes that confer resistance to autoimmune killing. This protocol can be applied in other mouse models of autoimmunity. For complete details on the use and execution of this protocol, please refer to Cai et al. (2020).(1)
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spelling pubmed-100252632023-03-21 Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model Li, Jian Lee, Yu-Chi Iessi, Isabela L. Wu, Chialing Yi, Peng Cai, Erica P. STAR Protoc Protocol Autoimmunity-induced pancreatic beta cell failure is the main characteristic of type 1 diabetes (T1D). Here, we describe a protocol for genome-scale in vivo CRISPR-Cas9 screening for use in a mouse model of T1D. Using a non-obese-diabetic-derived mouse beta cell line, NIT-1, and a genome-wide CRISPR-Cas9 knockout library (GeCKO-v2), we describe how to identify genes that confer resistance to autoimmune killing. This protocol can be applied in other mouse models of autoimmunity. For complete details on the use and execution of this protocol, please refer to Cai et al. (2020).(1) Elsevier 2023-03-13 /pmc/articles/PMC10025263/ /pubmed/36917606 http://dx.doi.org/10.1016/j.xpro.2023.102155 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Li, Jian
Lee, Yu-Chi
Iessi, Isabela L.
Wu, Chialing
Yi, Peng
Cai, Erica P.
Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title_full Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title_fullStr Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title_full_unstemmed Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title_short Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
title_sort protocol for genome-scale in vivo crispr screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025263/
https://www.ncbi.nlm.nih.gov/pubmed/36917606
http://dx.doi.org/10.1016/j.xpro.2023.102155
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