转录激活物样效应核酸酶
线粒体DNA
生物
遗传学
基因组编辑
点突变
基因
基因组
效应器
突变体
基因组工程
基因靶向
突变
细胞生物学
作者
Joachim Forner,Dennis Kleinschmidt,Etienne H. Meyer,Axel Fischer,Robert Morbitzer,Thomas Lahaye,Mark Aurel Schöttler,Ralph Bock
出处
期刊:Nature plants
[Springer Nature]
日期:2022-03-17
卷期号:8 (3): 245-256
被引量:23
标识
DOI:10.1038/s41477-022-01108-y
摘要
The development of technologies for the genetic manipulation of mitochondrial genomes remains a major challenge. Here we report a method for the targeted introduction of mutations into plant mitochondrial DNA (mtDNA) that we refer to as transcription activator-like effector nuclease (TALEN) gene-drive mutagenesis (GDM), or TALEN-GDM. The method combines TALEN-induced site-specific cleavage of the mtDNA with selection for mutations that confer resistance to the TALEN cut. Applying TALEN-GDM to the tobacco mitochondrial nad9 gene, we isolated a large set of mutants carrying single amino acid substitutions in the Nad9 protein. The mutants could be purified to homochondriomy and stably inherited their edited mtDNA in the expected maternal fashion. TALEN-GDM induces both transitions and transversions, and can access most nucleotide positions within the TALEN binding site. Our work provides an efficient method for targeted mitochondrial genome editing that produces genetically stable, homochondriomic and fertile plants with specific point mutations in their mtDNA.
科研通智能强力驱动
Strongly Powered by AbleSci AI