Generating an organ-deficient animal model using a multi-targeted CRISPR-Cas9 system.
Sci Rep
; 14(1): 10636, 2024 05 09.
Article
in En
| MEDLINE
| ID: mdl-38724644
ABSTRACT
Gene-knockout animal models with organ-deficient phenotypes used for blastocyst complementation are generally not viable. Animals need to be maintained as heterozygous mutants, and homozygous mutant embryos yield only one-fourth of all embryos. In this study, we generated organ-deficient embryos using the CRISPR-Cas9-sgRNAms system that induces cell death with a single-guide RNA (sgRNAms) targeting multiple sites in the genome. The Cas9-sgRNAms system interrupted cell proliferation and induced cell ablation in vitro. The mouse model had Cas9 driven by the Foxn1 promoter with a ubiquitous expression cassette of sgRNAms at the Rosa26 locus (Foxn1Cas9; Rosa26_ms). It showed an athymic phenotype similar to that of nude mice but was not hairless. Eventually, a rat cell-derived thymus in an interspecies chimera was generated by blastocyst complementation of Foxn1Cas9; Rosa26_ms mouse embryos with rat embryonic stem cells. Theoretically, a half of the total embryos has the Cas9-sgRNAms system because Rosa26_ms could be maintained as homozygous.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Forkhead Transcription Factors
/
CRISPR-Cas Systems
/
RNA, Guide, CRISPR-Cas Systems
Limits:
Animals
Language:
En
Journal:
Sci Rep
Year:
2024
Document type:
Article
Affiliation country:
Japan
Country of publication:
United kingdom