Purification and characterization of ribonucleoprotein effector complexes of Sulfolobus islandicus CRISPR-Cas systems.

Archaea CRISPR-Cas Systems Ribonucleoproteins / genetics Sulfolobus / genetics
His-tagged proteins Protein purification from Sulfolobales RNA-activated ssDNA cleavage assay Ribonucleoprotein complexes Target RNA cleavage assay crRNA

Journal

Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271

Informations de publication

Date de publication:
2021
Historique:
entrez: 9 11 2021
pubmed: 10 11 2021
medline: 18 3 2022
Statut: ppublish

Résumé

Archaea are preferred hosts for CRISPR-Cas systems. This adaptive immune system is not only widespread in archaeal organisms, but different types of CRISPR-Cas also co-exist in the same organism. Sulfolobus islandicus provides a good model for CRISPR research as genetic assays have been developed for revealing CRISPR immunity for the crenarchaeal model, and native ribonucleoprotein effector complexes have been expressed in this crenarchaeon and purified for characterization. Here we report a detailed protocol of purification and characterization of the Sulfolobus islandicus Cmr-β, the largest CRISPR effector known to date. The method can readily be applied to the purification of effectors encoded by other CRISPR-Cas systems in this organism, with the possibility to extend the application to other Sulfolobales.

Identifiants

DOI: 10.1016/bs.mie.2021.05.007 PMID: 34752293
pubmed: 34752293
pii: S0076-6879(21)00189-0
doi: 10.1016/bs.mie.2021.05.007
pii:
doi:

Substances chimiques

Ribonucleoproteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

327-347

Informations de copyright

Copyright © 2021 Elsevier Inc. All rights reserved.

Auteurs

Mingxia Feng (M)

CRISPR and Archaea Biology Research Center, Microbial Technology Institute and State Key Laboratory of Microbial Technology, Shandong University, Qingdao, PR, China.

Qunxin She (Q)

CRISPR and Archaea Biology Research Center, Microbial Technology Institute and State Key Laboratory of Microbial Technology, Shandong University, Qingdao, PR, China. Electronic address: shequnxin@sdu.edu.cn.

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Classifications MeSH

questionsmedicales.fr › Archéobactéries › Purification and characterization of...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › Purification and characterization of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Nucléoprotéines › Protéines de liaison à l'ARN › Ribonucléoprotéines › Purification and characterization of...
questionsmedicales.fr › Archéobactéries › Crenarchaeota › Sulfolobale › Sulfolobaceae › Sulfolobus › Purification and characterization of...