shRNA inhibits the expression of chicken telomerase reverse transcriptase in MDCC-MSB1 cells.
Genet Mol Res
; 15(1)2016 Jan 29.
Article
em En
| MEDLINE
| ID: mdl-26909930
ABSTRACT
Here, we investigated the effects of blocking chicken telomerase reverse transcriptase (chTERT) in MDCC-MSB1 cells, using small-hairpin RNAs (shRNAs) to interfere with gene expression. shRNAs specific to chTERT mRNA were designed, cloned into DNA plasmid vectors, and transfected into MDCC-MSB1 cells. The transfected chTERT RNAs were expressed by the RNA polymerase machinery of the MDCC-MSB1 cells. mRNA expression in transfected MDCC-MSB1 cells was detected using real-time PCR. After transfection, telomerase activity was monitored via a modified telomeric repeat amplification protocol assay, and cell cycle analysis was performed using a flow cytometer. At 72 h after transfection, chTERT expression was considerably reduced in cells transfected with shRNA; the highest inhibition rate was 89%. Compared with the control group, telomerase activity was significantly reduced and the cells failed to progress to S phase. shRNA effectively reduced telomerase activity and prohibited the transition of MDCC-MSB1 cells from G2/M to S phase.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Galinhas
/
Telomerase
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RNA Interferente Pequeno
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Interferência de RNA
Limite:
Animals
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article