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- W2954572711 endingPage "e3000350" @default.
- W2954572711 startingPage "e3000350" @default.
- W2954572711 abstract "Mutagenic screening is powerful for identifying key genes involved in developmental processes. However, such screens are successful only in lower organisms. Here, we develop a targeted genetic screening approach in mice through combining androgenetic haploid embryonic stem cells (AG-haESCs) and clustered regularly interspaced palindromic repeats/CRISPR-associated protein 9 (CRISPR-Cas9) technology. We produced a mutant semi-cloned (SC) mice pool by oocyte injection of AG-haESCs carrying constitutively expressed Cas9 and an single guide RNA (sgRNA) library targeting 72 preselected genes in one step and screened for bone-development–related genes through skeletal analysis at birth. This yielded 4 genes: Zic1 and Clec11a, which are required for bone development, and Rln1 and Irx5, which had not been previously considered. Whereas Rln1−/− mice exhibited small skeletal size only at birth, Irx5−/− mice showed skeletal abnormalities both in postnatal and adult phases due to decreased bone mass and increased bone marrow adipogenesis. Mechanistically, iroquois homeobox 5 (IRX5) promotes osteoblastogenesis and inhibits adipogenesis by suppressing peroxisome proliferator activated receptor γ (PPARγ) activation. Thus, AG-haESC-mediated functional mutagenic screening opens new avenues for genetic interrogation of developmental processes in mice." @default.
- W2954572711 created "2019-07-12" @default.
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- W2954572711 date "2019-07-02" @default.
- W2954572711 modified "2023-10-16" @default.
- W2954572711 title "Targeted genetic screening in mice through haploid embryonic stem cells identifies critical genes in bone development" @default.
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- W2954572711 doi "https://doi.org/10.1371/journal.pbio.3000350" @default.
- W2954572711 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6629148" @default.
- W2954572711 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/31265461" @default.
- W2954572711 hasPublicationYear "2019" @default.
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