論文 - 詳細
| RRC ID | 77291 |
|---|---|
| 著者 | Boddu PC, Gupta AK, Kim JS, Neugebauer KM, Waldman T, Pillai MM. |
| タイトル | Generation of scalable cancer models by combining AAV-intron-trap, CRISPR/Cas9, and inducible Cre-recombinase. |
| ジャーナル | Commun Biol |
| Abstract |
Scalable isogenic models of cancer-associated mutations are critical to studying dysregulated gene function. Nonsynonymous mutations of splicing factors, which typically affect one allele, are common in many cancers, but paradoxically confer growth disadvantage to cell lines, making their generation and expansion challenging. Here, we combine AAV-intron trap, CRISPR/Cas9, and inducible Cre-recombinase systems to achieve >90% efficiency to introduce the oncogenic K700E mutation in SF3B1, a splicing factor commonly mutated in multiple cancers. The intron-trap design of AAV vector limits editing to one allele. CRISPR/Cas9-induced double stranded DNA breaks direct homologous recombination to the desired genomic locus. Inducible Cre-recombinase allows for the expansion of cells prior to loxp excision and expression of the mutant allele. Importantly, AAV or CRISPR/Cas9 alone results in much lower editing efficiency and the edited cells do not expand due to toxicity of SF3B1-K700E. Our approach can be readily adapted to generate scalable isogenic systems where mutant oncogenes confer a growth disadvantage. |
| 巻・号 | 4(1) |
| ページ | 1184 |
| 公開日 | 2021-10-13 |
| DOI | 10.1038/s42003-021-02690-1 |
| PII | 10.1038/s42003-021-02690-1 |
| PMID | 34645977 |
| PMC | PMC8514589 |
| MeSH | CRISPR-Cas Systems / physiology* DNA Breaks, Double-Stranded Dependovirus Homologous Recombination Humans Integrases / physiology* Introns / physiology* Neoplasms / enzymology Neoplasms / genetics Neoplasms / physiopathology* |
| IF | 4.165 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 5 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HUDEP-2(RCB4557) |