Abstract：
:Tyrosinase is the rate-limiting enzyme for the production of melanin pigmentation. In the mouse and other animals, homozygous null mutations in the Tyrosinase gene (Tyr) result in the absence of pigmentation, i.e. albinism. Here we used the CRISPR/Cas9 system to generate mono- and bi-allelic null mutations in the Tyr locus by zygote injection of two single-guide and Cas9 RNAs. Injection into C57BL/6N wild-type embryos resulted in one completely albino founder carrying two different Tyr mutations. In addition, three pigmentation mosaics and fully pigmented littermates were obtained that transmitted new mutant Tyr alleles to progeny in test crosses with albinos. Injection into Tyr heterozygous (B6CBAF1/J×FVB/NJ) zygotes resulted in the generation of numerous albinos and also mice with a graded range of albino mosaicism. Deep sequencing revealed that the majority of the albinos and the mosaics had more than two new mutant alleles. These visual phenotypes and molecular genotypes highlight the somatic mosaicism and allele complexity in founders that occurs for targeted genes during CRISPR/Cas9-mediated mutagenesis by zygote injection in mice.
journal_name
Dev Bioljournal_title
Developmental biologyauthors
Yen ST,Zhang M,Deng JM,Usman SJ,Smith CN,Parker-Thornburg J,Swinton PG,Martin JF,Behringer RRdoi
10.1016/j.ydbio.2014.06.017subject
Has Abstractpub_date
2014-09-01 00:00:00pages
3-9issue
1eissn
0012-1606issn
1095-564Xpii
S0012-1606(14)00313-3journal_volume
393pub_type
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