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Journal of Southern Medical University ; (12): 1125-1130, 2021.
Artigo em Chinês | WPRIM | ID: wpr-888696

RESUMO

OBJECTIVE@#To explore the role of small nuclear noncoding RNA 7SK in embryonic stem cell (ESCs) proliferation and the value of 7SK as a target for early diagnosis and treatment for primordial dwarfism (PD).@*METHODS@#ESC line R1 was transfected with the CRISPR/Cas9 system, and sequencing of the PCR product and glycerol gradient analysis were performed to identify novel 7SK deletion mutations. A lentivirus system was used to knock down cyclin-dependent kinase 9 (CDK9) in clones with 7SK deletion mutations, and the effect of CDK9 knockdown on the protein level of cell division cycle 6 (CDC6) was analyzed with Western blotting.@*RESULTS@#We identified a novel deletion mutation of 7SK at 128-179 nt in the ESCs, which resulted in deficiency of cell proliferation. 7SK truncation at 128-179 nt significantly reduced the protein expressions of La-related protein 7 (LARP7) and CDC6.@*CONCLUSIONS@#7SK truncation at 128-179 nt can significantly impair proliferation of ESCs by downregulating CDC6. 7SK is a key regulator of proliferation and mediates the growth of ESCs through a mechanism dependent on CDK9 activity, suggesting the value of 7SK truncation at 128-179 nt as a potential target for early diagnosis and treatment of PD.


Assuntos
Humanos , Proteínas de Ciclo Celular , Proliferação de Células , Células-Tronco Embrionárias/metabolismo , Células HeLa , Proteínas Nucleares , Fator B de Elongação Transcricional Positiva/metabolismo , RNA Longo não Codificante/genética , Proteínas de Ligação a RNA , Ribonucleoproteínas , Fatores de Transcrição
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