論文 - 詳細
| RRC ID | 66881 |
|---|---|
| 著者 | You Y, Ramachandra SG, Jin T. |
| タイトル | A CRISPR-based method for testing the essentiality of a gene. |
| ジャーナル | Sci Rep |
| Abstract |
The CRISPR/Cas9 system is a powerful method of editing genes by randomly introducing errors into the target sites. Here, we describe a CRISPR-based test for gene essentiality (CRISPR-E test) that allows the identification of essential genes. Specifically, we use sgRNA-mediated CRISPR/Cas9 to target the open reading frame of a gene in the genome and analyze the in-frame (3n) and frameshift (3n + 1 and 3n + 2) mutations in the targeted region of the gene in surviving cells. If the gene is non-essential, the cells would carry both in-frame (3n) and frameshift (3n + 1 and 3n + 2) mutations. In contrast, the cells would carry only in-frame (3n) mutations if the targeted gene is essential, and this selective elimination of frameshift (3n + 1 and 3n + 2) mutations of the gene indicate its essentiality. As a proof of concept, we have used this CRISPR-E test in the model organism Dictyostelium discoideum to demonstrate that Dync1li1 is an essential gene while KIF1A and fAR1 are not. We further propose a simple method for quantifying the essentiality of a gene using the CRISPR-E test. |
| 巻・号 | 10(1) |
| ページ | 14779 |
| 公開日 | 2020-9-8 |
| DOI | 10.1038/s41598-020-71690-8 |
| PII | 10.1038/s41598-020-71690-8 |
| PMID | 32901070 |
| PMC | PMC7478968 |
| MeSH | Amino Acid Sequence Base Sequence CRISPR-Cas Systems* Dictyostelium / genetics* Dictyostelium / growth & development Dictyostelium / metabolism Gene Editing* Genes, Essential* Protozoan Proteins / antagonists & inhibitors Protozoan Proteins / genetics* Protozoan Proteins / metabolism Sequence Homology |
| IF | 3.998 |
| 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 | 10 |
| 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 | |
| 細胞性粘菌 | pTM1285 |