Novel off-Targeting Events Identified after Genome Wide Analysis of CRISPR-Cas Edited Pigs.

IF 3.7 4区 生物学 Q2 GENETICS & HEREDITY
CRISPR Journal Pub Date : 2024-06-01 Epub Date: 2024-05-21 DOI:10.1089/crispr.2024.0012
Bethany K Redel, Junchul Yoon, Emily Reese, Hong An, Kyungjun Uh, Paula R Chen, Randall S Prather, Kiho Lee
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引用次数: 0

Abstract

CRISPR-Cas technology has transformed our ability to introduce targeted modifications, allowing unconventional animal models such as pigs to model human diseases and improve its value for food production. The main concern with using the technology is the possibility of introducing unwanted modifications in the genome. In this study, we illustrate a pipeline to comprehensively identify off-targeting events on a global scale in the genome of three different gene-edited pig models. Whole genome sequencing paired with an off-targeting prediction software tool filtered off-targeting events amongst natural variations present in gene-edited pigs. This pipeline confirmed two known off-targeting events in IGH knockout pigs, AR and RBFOX1, and identified other presumably off-targeted loci. Independent validation of the off-targeting events using other gene-edited DNA confirmed two novel off-targeting events in RAG2/IL2RG knockout pig models. This unique strategy offers a novel tool to detect off-targeting events in genetically heterogeneous species after genome editing.

对 CRISPR-Cas 编辑过的猪进行全基因组分析后发现的新的非靶向事件。
CRISPR-Cas 技术改变了我们引入靶向修饰的能力,使猪等非常规动物模型能够模拟人类疾病,并提高其在食品生产中的价值。使用该技术的主要顾虑是可能会在基因组中引入不必要的修饰。在本研究中,我们展示了一种在三种不同基因编辑猪模型的基因组中全面识别全球范围内非靶向事件的方法。全基因组测序与脱靶预测软件工具相配合,在基因编辑猪的自然变异中筛选出脱靶事件。该管道确认了 IGH 基因敲除猪(AR 和 RBFOX1)中两个已知的非靶向事件,并确定了其他可能的非靶向位点。利用其他基因编辑 DNA 对脱靶事件进行的独立验证证实了 RAG2/IL2RG 基因敲除猪模型中的两个新的脱靶事件。这种独特的策略为检测基因组编辑后基因异质性物种的脱靶事件提供了一种新工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CRISPR Journal
CRISPR Journal Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.30
自引率
2.70%
发文量
76
期刊介绍: In recognition of this extraordinary scientific and technological era, Mary Ann Liebert, Inc., publishers recently announced the creation of The CRISPR Journal -- an international, multidisciplinary peer-reviewed journal publishing outstanding research on the myriad applications and underlying technology of CRISPR. Debuting in 2018, The CRISPR Journal will be published online and in print with flexible open access options, providing a high-profile venue for groundbreaking research, as well as lively and provocative commentary, analysis, and debate. The CRISPR Journal adds an exciting and dynamic component to the Mary Ann Liebert, Inc. portfolio, which includes GEN (Genetic Engineering & Biotechnology News) and more than 80 leading peer-reviewed journals.
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