ヒトのT細胞における全ゲノムCRISPRスクリーンは,免疫機能の重要な調節因子を明らかにする
Eric Shifrut1, Julia Carnevale2, Victoria Tobin1
1Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA; Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA 94720, USA.
Cell
|November 20, 2018
まとめ
私たちはSLICEを開発しました CRISPRによる 人間のT細胞のスクリーニングです このアプローチはT細胞の反応を制御する 重要な遺伝子を特定し 癌の免疫療法と薬剤発見に 極めて重要です
科学分野:
- 免疫学
- 分子生物学
- 遺伝学
背景:
- ヒトのT細胞は免疫と癌の免疫療法に不可欠です
- ヒトのT細胞におけるCRISPRスクリーンは 課題ですが 標的の発見には不可欠です
- T細胞刺激のレギュレータを特定することは 治療の進歩の鍵です
研究 の 目的:
- 原始的なヒトT細胞の新型CRISPRスクリーニング方法を開発する.
- T細胞受容体のシグナル伝達,増殖,抗がん活性を調節する遺伝子を特定する.
- 腫瘍の微小環境における 免疫抑制媒介を調べる
主な方法:
- Cas9タンパク質電解 (SLICE) による単一ガイドRNA (sgRNA) レンチウイルス感染という方法を開発した.
- 主要なヒトT細胞における全ゲノム機能喪失スクリーニングを行った.
- 単細胞RNA配列解析 (RNA-seq) と併用してSLICEを使用した.
主要な成果:
- T細胞受容体の重要なシグナリング成分と増殖のネガティブレギュレータを特定した.
- 癌細胞の殺戮を助長する 遺伝子の変異を特徴づけた
- 免疫抑制のメカニズムであるアデノシンシグナルへの反応を制御する遺伝子を明らかにした.
結論:
- SLICEプラットフォームは,原始ヒトT細胞の効率的で大規模なCRISPRスクリーニングを可能にします.
- SLICEは,免疫療法と薬の開発のための機能的な遺伝子標的の偏見のない発見を容易にする.
- この方法はT細胞の生物学と 治療再プログラムに関する理解を深めるものです
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