Genome Biology, 2021-06, Vol.22 (1), p.170-11, Article 170
2021
Details
- Autor(en) / Beteiligte
- Titel
- Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor
- Ist Teil von
- Genome Biology, 2021-06, Vol.22 (1), p.170-11, Article 170
- Ort / Verlag
- England: BioMed Central
- Erscheinungsjahr
- 2021
- Link zum Volltext
- Quelle
- SpringerLink
- Beschreibungen/Notizen
- Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1474-760X, 1474-7596eISSN: 1474-760XDOI: 10.1186/s13059-021-02389-wTitel-ID: cdi_doaj_primary_oai_doaj_org_article_090e19d9eb3b4d668ab587bfea39465f
- Format
- –
- Schlagworte
- Adamts20, Animal models, Animals, Base Sequence, Binding sites, Cell Line, Cells - metabolism, Chromatin, Chromatin - metabolism, Conversion, CRISPR, Dwarf phenotype, Editors, Efficiency, Embryo, Mammalian - metabolism, Embryos, Gene Editing, Genomes, Genotype & phenotype, Germline transmission, Humans, Igf2, Insulin-like growth factor II, Mice, Mice, Inbred C57BL, Mice, Inbred ICR, Mouse cells and embryos, Mutagenesis, Mutagenesis - genetics, Mutation, Peptides, Phenotypes, Plasmids, Prime editor, RNA, Guide, CRISPR-Cas Systems - genetics, RNA-directed DNA polymerase, Short Report, Targeted mutagenesis