Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Methods in Molecular Biology Année : 2021

Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification

Résumé

The need to generate modified cell lines that express tagged proteins of interest has become increasingly important. Here, we describe a detailed protocol for facile CRISPR/Cas9-mediated gene tagging and isolation of modified cells. In this protocol, we combine two previously published strategies that promote CRISPR/Cas9-mediated gene tagging: using chemically modified single-stranded oligonucleotides as donor templates and a co-selection strategy targeting the ATP1A1 gene at the same time as the gene of interest. Altogether, the protocol proposed here is both easier and saves time compared to other approaches for generating cells that express tagged proteins of interest, which is crucial to purify native complex from human cells.

Domaines

Génétique
Fichier principal
Vignette du fichier
480352_3_En_8_Chapter_Author.pdf (653.03 Ko) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-03706664 , version 1 (22-11-2022)

Identifiants

Citer

Sylvain Geny, Simon Pichard, Arnaud Poterszman, Jean-Paul Concordet. Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification. Methods in Molecular Biology, 2021, 2305, pp.153-174. ⟨10.1007/978-1-0716-1406-8_8⟩. ⟨hal-03706664⟩
57 Consultations
89 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More