KMID : 0368420220650060435
|
|
Journal of Plant Biology 2022 Volume.65 No. 6 p.435 ~ p.443
|
|
Identification of CRISPR-Induced Mutations in Plants: with a Focus on the Next-Generation Sequencing Assay
|
|
Yun Jae-Young
Yu Si-In Bang Seung-Eun Kim Jae-Yeon Lee Sang-Ho Lee Byeong-Ha
|
|
Abstract
|
|
|
As the CRISPR/Cas system is becoming an indispensable and most effective gene editing tool in eukaryotic cells, the need for efficient analysis of the gene editing outcomes also grows. Notably, ¡®in planta¡¯ gene editing, carried out by the Cas/sgRNA transgene after Agrobacterium-mediated plant transformation, demands several effective strategies to detect CRISPR gene-edited mutations. It is because it lacks ¡°cell screening¡± steps which are commonly included in the CRISPR gene editing in most mammalian or plant protoplast systems. In this review, we summarized and compared several CRISPR-mutant screening assays to facilitate the identification of correctly edited alleles. We especially described the Next-Generation Sequencing (NGS)-based analysis in detail. We hope that many labs can employ this strategy as the NGS screening is precise and quantitative and can be applied to large-scale screening while still being quantitative and accurate.
|
|
KEYWORD
|
|
CRISPR, Gene editing, Next-generation sequencing, Genome editing, CRISPR-mutant screening
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|