Abstract
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.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antineoplastic Agents / pharmacology
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Antineoplastic Agents / therapeutic use
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CRISPR-Associated Protein 9 / genetics
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CRISPR-Cas Systems / genetics*
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Carcinoma, Non-Small-Cell Lung / drug therapy*
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Carcinoma, Non-Small-Cell Lung / genetics
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Cell Line, Tumor
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Doxycycline / pharmacology
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Drug Discovery / methods*
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Drug Screening Assays, Antitumor / methods
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Female
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Gene Editing / methods*
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Gene Expression / drug effects
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Gene Expression / genetics
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Gene Expression Regulation, Neoplastic / drug effects
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Genetic Vectors / genetics
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HEK293 Cells
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High-Throughput Screening Assays / methods
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Humans
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Lung Neoplasms / drug therapy*
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Lung Neoplasms / genetics
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Male
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Mice
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Mice, Transgenic
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RNA, Guide, CRISPR-Cas Systems / genetics
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Recombination, Genetic / drug effects
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Reproducibility of Results
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Transcriptional Activation / drug effects
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Transfection / methods
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Transgenes / genetics
Substances
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Antineoplastic Agents
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RNA, Guide, CRISPR-Cas Systems
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CRISPR-Associated Protein 9
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Cas9 endonuclease Streptococcus pyogenes
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Doxycycline