Targeted genome editing in Caenorhabditis elegans using CRISPR/Cas9.

摘要: Utilization of programmable nucleases to generate DNA lesions at precise endogenous sequences has transformed the ability edit genomes from microbes plants and animals. This is especially true in organisms that previously lacked means engineer genomic changes, like Caenorhabditis elegans. C. elegans a 1 mm long free-living, nonparasitic, nematode worm, which easily cultivated laboratory. Its detailed genetic map relatively compact genome (~100 megabases) helped make it first metazoan have its entire sequenced. With sequence information came development numerous molecular tools dissect gene function. Initially absent this toolbox, however, were methods edits chosen loci. Adapting site-specific for use elegans, revolutionized studies biology. Zinc-finger (ZFNs), transcription activator-like effector (TALENs), then CRISPR-associated protein 9 (Cas9) used target specific double-strand breaks (DSBs). Precise changes could be engineered by providing repair templates targeting DSB trans. The ease programming Cas9 bind cleave with few limitations led widespread research sped strategies facilitate mutant recovery. Numerous innovative CRISPR/Cas9 methodologies are now primed elegans. For further resources related article, please visit WIREs website.