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Detection of MicroRNAs in Microglia by Real-time PCR in Normal CNS and During Neuroinflammation

Detection of MicroRNAs in Microglia by Real-time PCR in Normal CNS and During Neuroinflammation

Published on: July 23, 2012

一个整合的人类微质功能基因组学工具包.

Imdadul Haq^1,2,3, Jason C Ngo^1,2,3, Nainika Roy^1,2,3

¹Center for Translational and Computational Neuroimmunology, Columbia University Medical Center, New York, NY, USA.

Stem cell research & therapy

|April 10, 2024

概括

此摘要是机器生成的。

我们开发了一个快速的20天的协议,以产生真实的人类诱导的多能干细胞衍生微质细胞 (iMG) 用于阿尔茨海默病研究. 这个工具包使功能基因组学研究和神经退行性疾病中的遗传变异的调查成为可能.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?染色体的可访问性 (ATAC-Seq)功能性基因组学是一种功能性基因组学.微质细胞中的微质细胞神经退行性疾病的神经退行性疾病蛋白质组学是指蛋白质组学.由iPSC衍生的微质细胞 (iMG)

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科学领域:

神经科学是一个神经科学.
免疫学免疫学免疫学
遗传学是一个遗传学.

背景情况:

微质细胞是关键的大脑免疫细胞,参与发育和阿尔茨海默氏症 (AD) 等疾病.
人类诱导的多能干细胞衍生微质细胞 (iMG) 是有价值的模型,但目前的协议很长,缺乏全面的表征.
现有的方法限制了使用CRISPR-Cas9在iMG中的基因功能研究.

研究的目的:

开发一个优化,快速的协议,用于生成高质量的iMG.
在iMG中创建功能基因组学研究工具包.
促进研究神经退行性疾病中的遗传变异.

主要方法:

一个简化的两步,20天的协议,用于iPSC分化成iMG与正常的型.
使用单细胞RNA测序,ATAC-Seq,蛋白质组学和功能分析进行表征.
整合一种药物依赖的CRISPR-ON/OFF系统来控制基因表达和一个在线的多原子数据平台.

主要成果:

生成的iMG在转录,蛋白质和表观遗传方面与人类初级微细胞密切相似.
已证明iMG的功能,包括Ca2+过渡物,细胞因子反应,迁移和AD相关基质的细胞化.
确立了iMG作为适合在监管区域研究AD相关单核酸变异 (SNV) 的模型.

结论:

开发的协议快速有效地产生真实的iMG.
该工具包支持强大的微质功能基因组学和SNV调查.
这一进步有望在神经退行性疾病药物发现和治疗方面取得进展.