Development of an ObLiGaRe Doxycycline Inducible Cas9 system for pre-clinical cancer drug discovery.

Animals Antineoplastic Agents / pharmacology CRISPR-Associated Protein 9 / genetics CRISPR-Cas Systems / genetics Carcinoma, Non-Small-Cell Lung / drug therapy Cell Line, Tumor Doxycycline / pharmacology Drug Discovery / methods Drug Screening Assays, Antitumor / methods Female Gene Editing / methods Gene Expression / drug effects Gene Expression Regulation, Neoplastic / drug effects Genetic Vectors / genetics HEK293 Cells High-Throughput Screening Assays / methods Humans Lung Neoplasms / drug therapy Male Mice Mice, Transgenic RNA, Guide, Kinetoplastida / genetics Recombination, Genetic / drug effects Reproducibility of Results Transcriptional Activation / drug effects Transfection / methods Transgenes / genetics

Journal

Nature communications

ISSN: 2041-1723

Titre abrégé: Nat Commun

Pays: England

ID NLM: 101528555

Informations de publication

Date de publication:
29 09 2020

Historique:

received: 23 03 2020

accepted: 26 08 2020

entrez: 30 9 2020

pubmed: 1 10 2020

medline: 21 10 2020

Statut: epublish

Résumé

The CRISPR-Cas9 system has increased the speed and precision of genetic editing in cells and animals. However, model generation for drug development is still expensive and time-consuming, demanding more target flexibility and faster turnaround times with high reproducibility. The generation of a tightly controlled ObLiGaRe doxycycline inducible SpCas9 (ODInCas9) transgene and its use in targeted ObLiGaRe results in functional integration into both human and mouse cells culminating in the generation of the ODInCas9 mouse. Genomic editing can be performed in cells of various tissue origins without any detectable gene editing in the absence of doxycycline. Somatic in vivo editing can model non-small cell lung cancer (NSCLC) adenocarcinomas, enabling treatment studies to validate the efficacy of candidate drugs. The ODInCas9 mouse allows robust and tunable genome editing granting flexibility, speed and uniformity at less cost, leading to high throughput and practical preclinical in vivo therapeutic testing.

Identifiants

DOI: 10.1038/s41467-020-18548-9 PMID: 32994412 PMC: PMC7525522

pubmed: 32994412

doi: 10.1038/s41467-020-18548-9

pii: 10.1038/s41467-020-18548-9

pmc: PMC7525522

doi:

Substances chimiques

Antineoplastic Agents 0

RNA, Guide 0

CRISPR-Associated Protein 9 EC 3.1.-

Cas9 endonuclease Streptococcus pyogenes EC 3.1.-

Doxycycline N12000U13O

Types de publication

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

Langues

eng

Sous-ensembles de citation

Pagination

4903

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