title: shRNA inhibits the expression of chicken telomerase reverse transcriptase in MDCC-MSB1 cells.
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.
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun, Jilin, China.
authors.forename: N
authors.initials: N
authors.lastname: Jiang
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun, Jilin, China.
authors.forename: X B
authors.initials: XB
authors.lastname: Zheng
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun, Jilin, China.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Zhao
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun, Jilin, China.
authors.forename: Z G
authors.initials: ZG
authors.lastname: Qin
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun, Jilin, China.
authors.forename: T J
authors.initials: TJ
authors.lastname: Liu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002645:Chickens / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; D019098:Telomerase