Inhibiting GDF-8 expression by retrovirus-based RNAi stably / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 250-255, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-276131
ABSTRACT
We cloned human U6 promoter from pAVU6 + 27 vector into pXSN to transcripe small RNA. Meanwhile, a shRNA targeting GDF-8 was cloned down-stream of the hU6 promoter to construct recombinant vector. Then the packing cell GP-293 was co-transfected the recombinant with pVSV-G to gernarate virus particle. Resistant C2C12 cell pools were screened using G418. Levels of mRNA and protein of GDF-8 were tested by Real-Time PCR and western blotting. Cell proliferation and cell cycle were analyzed using MTT and FACS. The expression of GDF-8 was dramatically decreased by the retrovirus-based system in C2C12 cells. Cells proliferated effectively after integrating the recombinant. The cells in G0/G1 phase decreased by 13.7%, while cells in S phase increased by 14.9%. In conclusion, the retrovirus-based RNAi could be used to stably silence GDF-8. It can be a powerful tool in curing muscle atrophy.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Retroviridae
/
RNA Mensageiro
/
Regulação para Baixo
/
Células Cultivadas
/
Regulação da Expressão Gênica
/
Biologia Celular
/
Mioblastos
/
RNA Interferente Pequeno
/
Interferência de RNA
/
Proliferação de Células
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2008
Tipo de documento:
Artigo
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