相关实验视频

Updated: Jul 24, 2025

In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity

In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity

Published on: November 2, 2020

通过AAV插曲序列 (CrAAVe-seq) 进行CRISPR查是一种高度可扩展的细胞类型特定的in vivo查平台

Biswarathan Ramani^1,2, Indigo V L Rose^1,3, Noam Teyssier^1,4

¹Institute for Neurodegenerative Diseases; Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, USA.

bioRxiv : the preprint server for biology

|July 3, 2023

概括

我们开发了CRAAVe-seq,这是一个使用腺相关病毒 (AAV) 载体的CRISPR查平台,用于小鼠组织中细胞类型特定的遗传查. 这种方法有效地识别了目标细胞群中的必需基因,如神经元.

科学领域:

遗传学遗传学是一个
分子生物学分子生物学
神经科学是一个神经科学.

背景情况:

基于CRISPR的基因查需要可扩展的,体内*的方法来对哺乳动物组织进行细胞类型特定的分析.
现有的方法往往缺乏复杂组织环境所需的特异性和可扩展性.

研究的目的:

开发和验证基于腺相关病毒 (AAV) 的CRISPR查平台 (CrAAVe-seq),用于小鼠组织中细胞类型特定的遗传查.
通过大规模的聚合查,对神经元至关重要的基因进行分析 *in vivo*.
为优化*in vivo*CRISPR屏幕制定指导方针.

主要方法:

开发了CRAAVe-seq,这是一个基于AAV的平台,具有Cre敏感的sgRNA结构,用于在目标细胞群中进行聚合查.
在小鼠大脑中选了两个大型sgRNA库 (准>5,000个基因).
在初级神经元和*in vivo*中验证了确定的神经元必需基因.

主要成果:

在小鼠大脑中生成了神经元必需基因的强大概况.
验证了两个被确定为神经元功能必不可少的基因,无论是在体外*还是体内*.
证明了平台的可复制性,可扩展性和灵敏性,即使对于小的神经元亚群.

结论:

CrAAVe-seq是一种强大的,可扩展的工具,用于哺乳动物组织中细胞类型特定的*in vivo*遗传查.
该平台准确地识别了必要的基因,并为优化大规模屏幕提供了指导方针.
这种方法推进了复杂生物系统中基因功能的研究.

更多相关视频

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants

Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus AAV Capsid Variants

Published on: October 18, 2022

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

Published on: March 31, 2022

相关概念视频

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...