清脆的
生物
基因组编辑
Cas9
遗传学
亚基因组mRNA
苜蓿
植物抗病性
计算生物学
索引
基因
基因型
植物
单核苷酸多态性
作者
Udaya Subedi,Kimberley Burton Hughes,Guanqun Chen,Abdelali Hannoufa,Stacy D. Singer
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 219-239
被引量:1
标识
DOI:10.1007/978-1-0716-3159-1_16
摘要
CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) has become a breeding tool of choice for eliciting targeted genetic alterations in crop species as a means of improving a wide range of agronomic traits, including disease resistance, in recent years. With the recent development of CRISPR/Cas9 technology in Medicago sativa (alfalfa), which is an important perennial forage legume grown worldwide, its use for the enhancement of pathogen resistance is almost certainly on the horizon. In this chapter, we present detailed procedures for the generation of a single nonhomologous end-joining-derived indel at a precise genomic locus of alfalfa via CRISPR/Cas9. This method encompasses crucial steps in this process, including guide RNA design, binary CRISPR vector construction, Agrobacterium-mediated transformation of alfalfa explants, and molecular assessments of transformed genotypes for transgene and edit identification.
科研通智能强力驱动
Strongly Powered by AbleSci AI