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在体内大规模并行Perturb-seq显示皮层发育中的细胞类型特异性转录网络

Xinhe Zheng¹, Boli Wu¹, Yuejia Liu¹

¹Department of Neuroscience, Dorris Neuroscience Center, Scripps Research, La Jolla, CA 92037, USA.

|May 21, 2024

概括

这项研究开发了一个可扩展的体内CRISPR查平台,使用腺相关病毒 (AAV) 来进行基因传递和单细胞转录组. 这种增强的方法显著提高了标记效率,使复杂生物系统中的基因功能能够进行更深入的分析.

科学领域:

神经科学
遗传学
分子生物学

背景情况:

腺相关病毒 (AAV) 为体内研究提供了多样化的热带性和标记能力.
目前的体内CRISPR查方法在规模和效率方面存在局限性.

研究的目的:

扩大使用AAV和单细胞转录基因表型的体内CRISPR查规模.
在体内加速基因传递并提高标记效率.

主要方法:

在各种血清型中对86个AAV载体进行了广泛的测试,并使用了转位子系统.
胚胎到成人大脑和周围神经系统的单细胞转录基因表型.
开发一个巨大的平行体内Perturb-seq平台.

主要成果:

从几周到几天的时间,
在第6层皮质体神经元的细胞命运规范中确定了Foxg1的性作用.
达到了>6%的脑细胞标记,超过了lentivirus的疗效 (<0.1%),使得每次实验可以分析>30,000个细胞.

结论:

开发的平台可以通过高通量单细胞分析进行大规模的体内CRISPR查.
这种方法在体内促进了多种细胞类型的基因功能质疑.
这个平台将基因变异转化为因果功能, 进步我们对生物系统的理解.

关键词:

AAV 载体在CRISPR屏幕上大脑发育皮质发生在体内Perturb-seq 单细胞基因组学

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相关概念视频

RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...