Protocol for nuclear export signal characterization of cGAS in mammalian cells

The cyclic GMP-AMP synthase (cGAS) is the principal DNA sensor, which binds DNA and triggers the type I interferon production. We used ISD45 or inactivated Vaccinia Virus (VACV) to stimulate cGAS and monitored cellular localization by immunofluorescence microscopy, Operetta high-content screening, a...

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Detalles Bibliográficos
Autores principales: Huang, Yu, McLean, Myles, Liang, Chen, Guo, Fei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8261000/
https://www.ncbi.nlm.nih.gov/pubmed/34278335
http://dx.doi.org/10.1016/j.xpro.2021.100649
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author Huang, Yu
McLean, Myles
Liang, Chen
Guo, Fei
author_facet Huang, Yu
McLean, Myles
Liang, Chen
Guo, Fei
author_sort Huang, Yu
collection PubMed
description The cyclic GMP-AMP synthase (cGAS) is the principal DNA sensor, which binds DNA and triggers the type I interferon production. We used ISD45 or inactivated Vaccinia Virus (VACV) to stimulate cGAS and monitored cellular localization by immunofluorescence microscopy, Operetta high-content screening, and cytoplasmic/nuclear fractionation. LocNES server was used to predict cGAS nuclear export signal (NES) sequence and characterized the function by mutagenesis. This protocol provides a prototype of cGAS subcellular distribution or the identification of NES in other proteins. For complete details on the use and execution of this protocol, please refer to Sun et al. Sun et al. (2021)
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spelling pubmed-82610002021-07-16 Protocol for nuclear export signal characterization of cGAS in mammalian cells Huang, Yu McLean, Myles Liang, Chen Guo, Fei STAR Protoc Protocol The cyclic GMP-AMP synthase (cGAS) is the principal DNA sensor, which binds DNA and triggers the type I interferon production. We used ISD45 or inactivated Vaccinia Virus (VACV) to stimulate cGAS and monitored cellular localization by immunofluorescence microscopy, Operetta high-content screening, and cytoplasmic/nuclear fractionation. LocNES server was used to predict cGAS nuclear export signal (NES) sequence and characterized the function by mutagenesis. This protocol provides a prototype of cGAS subcellular distribution or the identification of NES in other proteins. For complete details on the use and execution of this protocol, please refer to Sun et al. Sun et al. (2021) Elsevier 2021-07-03 /pmc/articles/PMC8261000/ /pubmed/34278335 http://dx.doi.org/10.1016/j.xpro.2021.100649 Text en © 2021 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
Huang, Yu
McLean, Myles
Liang, Chen
Guo, Fei
Protocol for nuclear export signal characterization of cGAS in mammalian cells
title Protocol for nuclear export signal characterization of cGAS in mammalian cells
title_full Protocol for nuclear export signal characterization of cGAS in mammalian cells
title_fullStr Protocol for nuclear export signal characterization of cGAS in mammalian cells
title_full_unstemmed Protocol for nuclear export signal characterization of cGAS in mammalian cells
title_short Protocol for nuclear export signal characterization of cGAS in mammalian cells
title_sort protocol for nuclear export signal characterization of cgas in mammalian cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8261000/
https://www.ncbi.nlm.nih.gov/pubmed/34278335
http://dx.doi.org/10.1016/j.xpro.2021.100649
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AT guofei protocolfornuclearexportsignalcharacterizationofcgasinmammaliancells

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