Construction and evaluation of gene silencing vectors of Noggin
Author:
Affiliation:

Clc Number:

Fund Project:

  • Article
  • |
  • Figures
  • |
  • Metrics
  • |
  • Reference
  • |
  • Related
  • |
  • Cited by
  • |
  • Materials
  • |
  • Comments
    Abstract:

    Objective To construct the retroviral-mediated short hairpin RNA(shRNA) expression vectors of Noggin, and to analyze the silencing effect of the shRNA. Methods Four interference sequences were designed based onNoggin mRNA, and connected to Lenti-KD vectors. The recombinant plasmids were transiently transfected into HEK-293T cells, and finally obtained the recombinant virus.Then MC3T3-E1 cells were infected with the recombinant virus and screened by puromycin.The gene silencing effect of shRNA was evaluated using RT-PCR and Western blot. Results RT-PCR results showed that all the four interference sequences had silencing effect on the expression of Noggin, but only shNoggin-1(P<0.01) was different significantly. Western blot results showed that among the four interference sequences, only shNoggin-1(P<0.01) showed a significantly lowering effect on the expression of Noggin. Conclusions One interference sequence of Noggin gene is obtained, which can interfere the stability of mRNA of the Noggin gene, by affecting the protein expression. It is useful for the further study of unknown biological function of Noggin.

    Reference
    Related
    Cited by
Get Citation
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:July 14,2015
  • Revised:
  • Adopted:
  • Online: March 05,2016
  • Published: