Protocol for nuclear export signal characterization of cGAS in mammalian cells
Summary: 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 scr...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2021-09-01
|
Series: | STAR Protocols |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721003567 |
id |
doaj-2b26fd1244f4474bbf2c3caebc1f071d |
---|---|
record_format |
Article |
spelling |
doaj-2b26fd1244f4474bbf2c3caebc1f071d2021-09-19T05:00:10ZengElsevierSTAR Protocols2666-16672021-09-0123100649Protocol for nuclear export signal characterization of cGAS in mammalian cellsYu Huang0Myles McLean1Chen Liang2Fei Guo3NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P. R. China; Corresponding authorMcGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, CanadaMcGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, CanadaNHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P. R. China; Corresponding authorSummary: 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)http://www.sciencedirect.com/science/article/pii/S2666166721003567Cell BiologyImmunologyMolecular Biology |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yu Huang Myles McLean Chen Liang Fei Guo |
spellingShingle |
Yu Huang Myles McLean Chen Liang Fei Guo Protocol for nuclear export signal characterization of cGAS in mammalian cells STAR Protocols Cell Biology Immunology Molecular Biology |
author_facet |
Yu Huang Myles McLean Chen Liang Fei Guo |
author_sort |
Yu Huang |
title |
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_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_sort |
protocol for nuclear export signal characterization of cgas in mammalian cells |
publisher |
Elsevier |
series |
STAR Protocols |
issn |
2666-1667 |
publishDate |
2021-09-01 |
description |
Summary: 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) |
topic |
Cell Biology Immunology Molecular Biology |
url |
http://www.sciencedirect.com/science/article/pii/S2666166721003567 |
work_keys_str_mv |
AT yuhuang protocolfornuclearexportsignalcharacterizationofcgasinmammaliancells AT mylesmclean protocolfornuclearexportsignalcharacterizationofcgasinmammaliancells AT chenliang protocolfornuclearexportsignalcharacterizationofcgasinmammaliancells AT feiguo protocolfornuclearexportsignalcharacterizationofcgasinmammaliancells |
_version_ |
1717376193366327296 |