In:
Development, The Company of Biologists
Abstract:
In vivo genetic mutations has become a powerful tool for dissecting gene function; however, multi-gene interaction and compensatory mechanisms involving can make findings from single mutations at best difficult to interpret, and at worst, misleading. Hence, it is necessary to establish an efficient way to disrupt multiple genes simultaneously. The CRISPR/Cas9-mediated base editing disrupts gene function by converting a protein coding sequence into a stop codon; this is referred to as CRISPR-stop. Its application in generating zygotic mutations has not been well explored yet. Here, we firstly performed a proof-of-principle test by disrupting Atoh1, a gene critical for auditory hair cell generation. Next, we individually mutated vGlut3, Otoferlin and Prestin, three genes needed for normal hearing function. Finally, we successfully disrupted vGlut3, Otoferlin and Prestin simultaneously. Our results show that CRISPR-stop can efficiently generate single or triple homozygous F0 mice mutants, bypassing laborious mouse breeding. We believe that CRISPR-stop is a powerful method that will pave the way for high-throughput screening mouse developmental and functional genes, matching the efficiency of methods available for model organisms, such as Drosophila.
Type of Medium:
Online Resource
ISSN:
1477-9129
,
0950-1991
Language:
English
Publisher:
The Company of Biologists
Publication Date:
2018
detail.hit.zdb_id:
2007916-3
SSG:
12
Permalink