論文 - 詳細
| RRC ID | 45911 |
|---|---|
| 著者 | Kage-Nakadai E, Imae R, Suehiro Y, Yoshina S, Hori S, Mitani S. |
| タイトル | A conditional knockout toolkit for Caenorhabditis elegans based on the Cre/loxP recombination. |
| ジャーナル | PLoS One |
| Abstract |
Conditional knockout (cKO) based on site-specific recombination (SSR) technology is a powerful approach for estimating gene functions in a spatially and temporally specific manner in many model animals. In Caenorhabditis elegans (C. elegans), spatial- and temporal-specific gene functions have been largely determined by mosaic analyses, rescue experiments and feeding RNAi methods. To develop a systematic and stable cKO system in C. elegans, we generated Cre recombinase expression vectors that are driven by various tissue-specific or heat-shock promoters. Validation using Cre-mediated fluorescence protein inactivation or activation systems demonstrated successful Cre-dependent loxP excision. We established a collection of multi-copy Cre transgenic strains for each evaluated vector. To evaluate our Cre/loxP-based cKO system, we generated sid-1 deletion mutants harboring floxed sid-1 single-copy integration (SCI) using ultraviolet trimethylpsoralen (UV/TMP) methods. sid-1 mutants that were rescued by the floxed sid-1 SCI were then crossed with the Pdpy-7::Cre strain for cKO in the hypodermis. The sid-1 cKO animals were resistant to bli-3 RNAi, which causes the Bli-phenotyple in the hypodermis, but they were sensitive to unc-22 RNAi, which leads to twitching of the body wall muscle. Our system, which is based on the combination of a transgenic Cre collection, pre-existing deletion mutants, and UV/TMP SCI methods, provided a systematic approach for cKO in C. elegans. |
| 巻・号 | 9(12) |
| ページ | e114680 |
| 公開日 | 2014-1-1 |
| DOI | 10.1371/journal.pone.0114680 |
| PII | PONE-D-14-40651 |
| PMID | 25474529 |
| PMC | PMC4256423 |
| MeSH | Animals Animals, Genetically Modified Caenorhabditis elegans / genetics* Gene Knockout Techniques / methods* Genetic Engineering Genetic Vectors Green Fluorescent Proteins / biosynthesis Green Fluorescent Proteins / genetics Integrases / genetics Organ Specificity RNA Interference |
| IF | 2.74 |
| 引用数 | 11 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| 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 | 4 |
| 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 | |
| 線虫 | tm2700 tm234 tm246 |