Repositorium
HRM-facilitated rapid identification and genotyping of mutations induced by CRISPR/Cas9 mutagenesis in rice
316
Journal Article / 2018
Li, Shan; Liu, Songmei; Liu, Yanhua; Lu, Haiping; Tan, Yuanyuan; Huang, Jianzhong; Wei, Pengcheng; Shu, Qing Yao
Abstract: The clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated (Cas) technology has recently emerged as a powerful genomic editing tool with great potential for crop breeding. However, commonly used protocols for screening of CRISPR/Cas9-induced mutations are laborious, timeconsuming, and costly. In the present study we examined the applicability of high resolution melting (HRM) analysis fast screening CRISPR/Cas9-induced mutations in T0 plants and subsequent genotyping of T1 plants derived from single T0 tive analysis demonstrated that HRM analysis could identify mutant T0 typed by HRM analysis using WT parents and T0 populations. Comparaplants carrying various types of mutation, including single nucleotide substitutions and short insertion/deletions, with false positive/negative rates of 0-2.78%. Furthermore, T1 plants could be correctly genoplants as controls. We hence recommend the adoption of HRM analysis in CRISPR/Cas9 mediated genetic studies and breeding in rice and other crop species.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
736 |
Qing Yao Shu China |
Oryza sativa |
CRISPR/Cas9 BEL1 |
herbicide tolerance SDN1 |
Basic research Basic research |
737 |
Qing Yao Shu China |
Oryza sativa |
CRISPR/Cas9 LCT1 |
Encodes a cadmium transporter SDN1 |
Basic research Basic research |