相关实验视频

Updated: Jul 16, 2025

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

Published on: January 3, 2015

在indel软件解决与体质CRISPR/Cas9瘤发生模型之间的差异

Qierra R Brockman^1,2, Amanda Scherer¹, Gavin R McGivney^1,3,4

¹Department of Internal Medicine, Carver College of Medicine, University of Iowa, 375 Newton Rd, 5206 MERF, Iowa City, IA, 52246, USA.

Scientific reports

|September 8, 2023

概括

在CRISPR/Cas9基因编辑分析软件平台之间存在显著差异. 这项研究揭示了在体内小鼠模型中indel检测的高可变性,影响了瘤研究.

科学领域:

基因组工程和分子生物学.
癌症研究,特别是瘤建模.
生物信息学和计算生物学.

背景情况:

克里斯普尔/卡斯9技术是体内基因组工程的强大工具.
对插入和删除 (indels) 的准确表征对于CRISPR/Cas9应用至关重要.
现有的英德尔分析软件还没有系统地比较体质 in vivo 小鼠模型.

研究的目的:

为了比较四个不同的indel分析软件平台的性能.
为了评估软件的可变性,从体质 in vivo 小鼠模型中分析CRISPR/Cas9诱导的内置.
评估软件选择对恶性外围神经膜瘤 (MPNST) 模型中的indel特征的影响.

主要方法:

在小鼠模型中利用CRISPR/Cas9介导的NF1基因编辑的测序数据.
在同一数据集上应用了四个不同的公开可用的indel分析软件平台.
直接比较每个平台报告的indel的数量,规模和频率.

主要成果:

在四个软件平台上观察到indel检测的显著变化.
报告的不一致性,在数量,大小和频率的indels. .
变异性对于较大的indels特别明显,在体内瘤模型中很常见.

结论:

选择的indel分析软件显著影响了对CRISPR/Cas9编辑结果的解释.
高可变性需要仔细选择适合特定实验环境的分析平台,特别是对于体内模型.
这些发现强调了在复杂的生物系统 (如瘤模型) 中进行独立分析的标准化或验证方法的需要.

更多相关视频

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Published on: August 28, 2018

相关概念视频

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...