Construction and identification of lentiviral vector encoding shRNA against Smad3

WANG Yan-zhou; LIANG Zhi-qing; LIU Xiao-fang; XU Hui-cheng

您当前的位置：

首页 >

文章列表页 >

Construction and identification of lentiviral vector encoding shRNA against Smad3

更新时间：2025-01-15

- Construction and identification of lentiviral vector encoding shRNA against Smad3
- Journal of Army Medical University Vol. 31, Issue 8, Pages: 701-702(2009)
- 作者机构：
  
  第三军医大学西南医院妇产科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R394.3;R373;R394-33
- Published：2009，
  
  Published Online：30 April 2009，
- 稿件说明：
移动端阅览
WANG YAN-ZHOU, LIANG ZHI-QING, LIU XIAO-FANG, et al. Construction and identification of lentiviral vector encoding shRNA against Smad3. [J]. Journal of army medical university, 2009, 31(8): 701-702.
DOI：

WANG YAN-ZHOU, LIANG ZHI-QING, LIU XIAO-FANG, et al. Construction and identification of lentiviral vector encoding shRNA against Smad3. [J]. Journal of army medical university, 2009, 31(8): 701-702. DOI：

摘要

目的构建Smad3基因RNAi慢病毒载体。方法针对已经筛选确定的Smad3基因RNAi有效靶序列

合成靶序列的Oligo DNA

退火形成双链DNA

与经

Age

Ⅰ和

Eco

RⅠ双酶切后的pGCSIL-GFP载体连接产生GC-shSmad3慢病毒载体

PCR筛选阳性克隆

测序鉴定。用GC-shSmad3、pHelper 1.0 载体和pHelper 2.0载体共转染包装细胞293T细胞

包装产生慢病毒

以293T细胞GFP蛋白的表达水平测定病毒滴度。结果 PCR 和测序证实

构建出了Smad3 shRNA的慢病毒载体GC-shSmad3。包装慢病毒

浓缩病毒悬液的滴度为3×10

TU/ml。结论成功构建Smad3基因RNAi慢病毒载体。

Abstract

Objective To construct a lentiviral vector expressing small-hairpin RNA (shRNA) targeting Smad3 gene. Methods The targeting sequence of Smad3 gene which can be effectively silenced in RNA inference was confirmed in our previous study. The cDNA containing both sense and antisense Oligo DNA fragments of the targeting sequence was designed

synthesized and cloned into the pGCSIL-GFP vector. The obtained lentiviral vector containing Smad3 shRNA was confirmed by PCR and sequencing. 293T cells were cotransfected with lentiviral vector GC-shSmad3

pHelper 1.0 and pHelper 2.0. The titer of virus was tested according to the expression level of GFP. Results PCR and DNA sequencing demonstra

ted that the constructed lentivirus vector GC-shSmad3 produced Smad3 shRNA. The titer of concentrated virus was 3×10

TU/ml. Conclusion The lentivirus RNAi vector targeting Smad3 is constructed successfully.

关键词

RNA干扰Smad3慢病毒

Keywords

RNA interferenceSmad3lentivirus

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Construction of recombinant lentivirol expression vector carrying human Caveolin-1 with RNA interference and its effect on invasiveness of SMMC7721 cells

Effects of the inhibition of Smad3 by short hairpin RNA on the expression of Smad7 in keloid fibroblasts

Construction of lentiviral RNAi vectors containing human smoothened gene and its inhibitory effect on proliferation and differentiation in human periodontal ligament stem cells

Construction of lentiviral vector for SPAG6 RNA interference and its influence on apoptosis of SKM-1 cells

Related Author

ZHANG Zhi-bo

CAI Lei

ZHENG Shu-guo

XIONG Yan

DONG Jia-hong

DAN Yang

SHEN Wei-min

GUO Lei

Related Institution

Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University

Department of Plastic Surgery, First Affiliated Hospital, Chongqing Medical University

Department of Stomatology, Southwest Hospital, Third Military Medical University

Department of Stomatology, Out-patient Department of Sichuan Military Command

Department of Stomatology, Deyang Second People’s Hospital

⁰