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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes
- Source :
- Developmental Biology. (1):3-9
- Publisher :
- Published by Elsevier Inc.
-
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.
- Subjects :
- Male
Zygote
Tyrosinase
Mutant
CRISPR-Associated Proteins
Mice
0302 clinical medicine
CRISPR-Associated Protein 9
Genotype
Genetics
Gene Editing
0303 health sciences
Monophenol Monooxygenase
Mosaicism
Pigmentation
High-Throughput Nucleotide Sequencing
Embryo
Female
RNA, Guide, Kinetoplastida
Genome editing
Albinism
Knockout
Locus (genetics)
Biology
Article
03 medical and health sciences
Bacterial Proteins
Melanocyte
Animals
RNA, Messenger
Allele
Gene
Molecular Biology
Alleles
030304 developmental biology
Melanins
Base Sequence
Gene targeting
Sequence Analysis, DNA
Cell Biology
Endonucleases
Molecular biology
Mice, Inbred C57BL
Mutagenesis
Mutation
CRISPR-Cas Systems
Mosaic
030217 neurology & neurosurgery
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 00121606
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Developmental Biology
- Accession number :
- edsair.doi.dedup.....9265c2b83fab4afed4ccc14f4284d2e4
- Full Text :
- https://doi.org/10.1016/j.ydbio.2014.06.017