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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...

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高复杂度条形编码狂犬病病毒用于使用单细胞和单核测序进行可扩展电路映射.

David Shin^1,2, Madeleine E Urbanek^1,2, H Hanh Larson²

¹Biomedical Sciences Graduate Program, University of California, San Francisco, CA, USA.

bioRxiv : the preprint server for biology

|December 23, 2024

概括

新的狂犬病病毒 (RV) 技术使神经元细胞类型的可扩展电路映射成为可能. 这种方法将分子身份与突触连接联系起来,进步我们对健康和疾病中的大脑电路的理解.

关键词:

有条码的狂犬病病毒条形码复杂度条形码复杂度条形码多样性条形码多样性大脑发育大脑的发育大脑的发展皮层皮层皮层人类人类人类人类人类人类人类一个单细胞RNA测序.单核RNA测序的一个核.亚板子板是指一个子板子板.

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科学领域:

神经科学是一个神经科学.
基因组学就是基因组学.
分子生物学分子生物学

背景情况:

单细胞基因组学已经推进了神经元细胞类型识别.
要了解功能性大脑电路,需要用于绘制单细胞连接的可扩展方法.
目前对分子定义细胞类型如何形成电路的理解是有限的.

研究的目的:

开发一种可扩展的协议,使用条形码狂犬病病毒 (RV) 来绘制神经元连接的地图.
通过单核RNA测序 (snRNA-seq) 实现连接信息的读取.
研究细胞类型特异性电路图案在发育中的人类大脑中的出现.

主要方法:

产生高度复杂的条形码狂犬病病毒 (RV) 用于并行电路跟踪.
将RV编码的条形码转录定位到核中进行snRNA-seq分析.
该协议在正在发育的人类大脑皮层的器官类型切片培养中应用.

主要成果:

来自数万个起始细胞的突触连接映射的可扩展协议的演示.
在中型人类大脑皮层中识别细胞类型特定的电路图案.
成功整合了带条码的RV跟踪与snRNA-seq用于电路分析.

结论:

开发的协议提供了一个可扩展的方法,用于绘制分子定义细胞类型的突触连接.
这项技术为健康和疾病状态的电路映射提供了一条前进的道路.
揭示了功能性大脑电路在发育过程中的出现.