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2.
Exp Mol Med ; 54(12): 2188-2199, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36494589

RESUMO

The generation of conditional alleles using CRISPR technology is still challenging. Here, we introduce a Short Conditional intrON (SCON, 189 bp) that enables the rapid generation of conditional alleles via one-step zygote injection. In this study, a total of 13 SCON mouse lines were successfully generated by 2 different laboratories. SCON has conditional intronic functions in various vertebrate species, and its target insertion is as simple as CRISPR/Cas9-mediated gene tagging.


Assuntos
Sistemas CRISPR-Cas , Zigoto , Camundongos , Animais , Sistemas CRISPR-Cas/genética , Íntrons/genética , Técnicas de Inativação de Genes
3.
Cell Stem Cell ; 29(5): 826-839.e9, 2022 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-35523142

RESUMO

Adult stem cells constantly react to local changes to ensure tissue homeostasis. In the main body of the stomach, chief cells produce digestive enzymes; however, upon injury, they undergo rapid proliferation for prompt tissue regeneration. Here, we identified p57Kip2 (p57) as a molecular switch for the reserve stem cell state of chief cells in mice. During homeostasis, p57 is constantly expressed in chief cells but rapidly diminishes after injury, followed by robust proliferation. Both single-cell RNA sequencing and dox-induced lineage tracing confirmed the sequential loss of p57 and activation of proliferation within the chief cell lineage. In corpus organoids, p57 overexpression induced a long-term reserve stem cell state, accompanied by altered niche requirements and a mature chief cell/secretory phenotype. Following the constitutive expression of p57 in vivo, chief cells showed an impaired injury response. Thus, p57 is a gatekeeper that imposes the reserve stem cell state of chief cells in homeostasis.


Assuntos
Celulas Principais Gástricas , Inibidor de Quinase Dependente de Ciclina p57/metabolismo , Animais , Linhagem da Célula , Celulas Principais Gástricas/metabolismo , Camundongos , Organoides , Células-Tronco , Estômago
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