Small RNA Transfection in Primary Human Th17 Cells by Next Generation Electroporation.
Small RNA Transfection in Primary Human Th17 Cells by Next Generation Electroporation.
复制标题
下一代电穿孔中原代人Th17细胞中的小RNA转染。
DOI：
10.3791/55546
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发表时间：
2017-04-13
期刊：
影响因子：
--
通讯作者：
Ansel KM
中科院分区：
文献类型：
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作者：
Montoya MM;Ansel KM
氖®新一代电穿孔是一种用小RNA转染人Th 17细胞以改变基因表达和细胞行为的有效方法。CD 4 + T细胞可以根据周围的细胞因子环境分化为效应T辅助细胞的几个亚群。Th 17细胞可以通过在极化细胞因子IL-1β、IL-6、IL-23和TGFβ的存在下激活初始CD 4 + T细胞而在体外从初始CD 4 + T细胞产生。Th 17细胞协调针对细胞外细菌和真菌的免疫，但它们的异常活性也与几种自身免疫性和炎性疾病有关。Th 17细胞由趋化因子受体CCR 6识别，并由其主转录因子RORγt和特征性效应细胞因子IL-17 A定义。Th 17细胞分化的优化培养条件有助于在受控环境中研究人T细胞生物学的机制。它们还通过RNA干扰或引入microRNA（miRNA）模拟物或抑制剂来研究特定基因和基因表达程序的重要性。该方案提供了使用氖®下一代电穿孔装置用小RNA瞬时转染分化中的原代人Th 17细胞的易于使用、可再现且高效的方法。
The Neon® next generation electroporation is an efficient method for transfecting human Th17 cells with small RNAs to alter gene expression and cell behavior. CD4+ T cells can differentiate into several subsets of effector T helper cells depending on the surrounding cytokine milieu. Th17 cells can be generated from naive CD4+ T cells in vitro by activating them in the presence of the polarizing cytokines IL-1β, IL-6, IL-23, and TGFβ. Th17 cells orchestrate immunity against extracellular bacteria and fungi, but their aberrant activity has also been associated with several autoimmune and inflammatory diseases. Th17 cells are identified by the chemokine receptor CCR6 and defined by their master transcription factor, RORγt, and characteristic effector cytokine, IL-17A. Optimized culture conditions for Th17 cell differentiation facilitate mechanistic studies of human T cell biology in a controlled environment. They also provide a setting for studying the importance of specific genes and gene expression programs through RNA interference or the introduction of microRNA (miRNA) mimics or inhibitors. This protocol provides an easy to use, reproducible, and highly efficient method for transient transfection of differentiating primary human Th17 cells with small RNAs using the Neon® next generation electroporation device.
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影响因子：
29.7
作者：
Zhu J;Yamane H;Paul WE
通讯作者：
Paul WE
DOI：
10.1084/jem.20070663
发表时间：
2007-08-06
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Annunziato F;Cosmi L;Santarlasci V;Maggi L;Liotta F;Mazzinghi B;Parente E;Filì L;Ferri S;Frosali F;Giudici F;Romagnani P;Parronchi P;Tonelli F;Maggi E;Romagnani S
通讯作者：
Romagnani S
影响因子：
4.3
作者：
Kim, Tae Kyung;Eberwine, James H.
通讯作者：
Eberwine, James H.
影响因子：
30.5
作者：
Du, Changsheng;Liu, Chang;Pei, Gang
通讯作者：
Pei, Gang
影响因子：
30.5
作者：
通讯作者：
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