Résumé
Studying the cellular function of microRNAs requires genetic strategies to generate their loss-of-function. Recently, a novel approach of targeted genomic deletion was proposed, which is based on induction of site-specific DNA cuts with Cas9/gRNA ribonucleoprotein complexes, combined with homologous recombination-dependent insertion of cassettes that contain sequences for Cre-lox or FLP-FRT systems. Here, we provide a technical report describing application of this CRISPR/Cas9-directed homologous recombination procedure to the generation of human tumor cells in which conditional knockout of an X-linked cluster of microRNAs (miR-105/miR-767) can be induced. We describe the successive steps of genetic engineering and cell clone selection that allowed us to generate cells with the expected genome editing.
langue originale | Anglais |
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journal | Gene Technology |
Volume | 2015 |
Numéro de publication | 4:124 |
Les DOIs | |
Etat de la publication | Publié - 20 juil. 2015 |