Afficher la couverture 'Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification | Geny, Sylvain' en grand format
/5

0 avis

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

Archive ouverte

Geny, Sylvain | Pichard, Simon | Poterszman, Arnaud | Concordet, Jean-Paul

Edité par CCSD ; Humana Press/Springer Imprint -

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.

Suggestions

Du même auteur

Tagging Proteins with Fluorescent Reporters Using the CRISPR/Cas9 System and Double-Stranded DNA Donors

Archive ouverte | Geny, Sylvain | CCSD

International audience. Macromolecular complexes govern the majority of biological processes and are of great biomedical relevance as factors that perturb interaction networks underlie a number of diseases, and inhi...

Functional interplay between TFIIH and KAT2A regulates higher-order chromatin structure and class II gene expression

Archive ouverte | Sandoz, Jérémy | CCSD

International audience. The TFIIH subunit XPB is involved in combined Xeroderma Pigmentosum and Cockayne syndrome (XP-B/CS). Our analyses reveal that XPB interacts functionally with KAT2A, a histone acetyltransferas...

A Time and Cost-Effective Pipeline for Expression Screening and Protein Production in Insect Cells Based on the HR-Bac Toolbox to Generate Recombinant Baculoviruses

Archive ouverte | Loup-Forest, Jules | CCSD

International audience. The baculovirus expression vector system (BEVS) is recognized as a powerful platform for producing challenging proteins and multiprotein complexes both in academia and industry. Since a bacul...

Chargement des enrichissements...