Application of CRISPR/cas9-Directed Homologous Recombination to the Generation of Human Tumor Cells with Conditional Knockout of an X-Linked MicroRNA Locus

Aurélie Van Tongelen, Axelle Loriot, Olivier De Backer, Charles De Smet

Résultats de recherche: Contribution à un journal/une revueArticle

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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 originaleAnglais
journalGene Technology
Volume2015
Numéro de publication4:124
Les DOIs
étatPublié - 20 juil. 2015

Empreinte digitale

Clustered Regularly Interspaced Short Palindromic Repeats
Homologous Recombination
MicroRNAs
Guide RNA
Ribonucleoproteins
Genetic Engineering
Neoplasms
Clone Cells
DNA

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abstract = "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.",
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Application of CRISPR/cas9-Directed Homologous Recombination to the Generation of Human Tumor Cells with Conditional Knockout of an X-Linked MicroRNA Locus. / Van Tongelen, Aurélie; Loriot, Axelle; De Backer, Olivier; De Smet, Charles.

Dans: Gene Technology, Vol 2015, Numéro 4:124, 20.07.2015.

Résultats de recherche: Contribution à un journal/une revueArticle

TY - JOUR

T1 - Application of CRISPR/cas9-Directed Homologous Recombination to the Generation of Human Tumor Cells with Conditional Knockout of an X-Linked MicroRNA Locus

AU - Van Tongelen, Aurélie

AU - Loriot, Axelle

AU - De Backer, Olivier

AU - De Smet, Charles

PY - 2015/7/20

Y1 - 2015/7/20

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