Dissertation > Excellent graduate degree dissertation topics show

Construction of Recombinant shRNA-espressing Vectors Targeting to Two Different Hepatic Fibrosis-related Cytokines

Author: LiuJiaQun
Tutor: ZhangJianJun
School: Luzhou Medical College
Course: Internal Medicine
Keywords: RNA interference Liver fibrosis Plasmid vector Connective tissue growth factor Tissue inhibitor of metalloproteinase factor-1
CLC: R575.2
Type: Master's thesis
Year: 2009
Downloads: 33
Quote: 0
Read: Download Dissertation

Abstract


Objective: connective tissue growth factor (CTGF) is a recently discovered an important cytokine promoting liver fibrosis , transforming growth factor- β ( transforming growth factorβ TGF- β ) signaling pathway downstream effects of media . CTGF can directly mediated activation of primary hepatic stellate cells (hepatic stellate cell, HSC) proliferation and migration , but also promote the activation of HSC synthesis and secretion of extracellular matrix ( extracellular matrix , ECM) . Metal tissue inhibitor of metalloproteinase 1 (TIMP-1) of TGF-β signaling pathways downstream effects of media , liver fibrosis , which is mainly expressed by activated HSC . TIMP-1 not only inhibits the activity of matrix metalloproteinases (matrix metalloproteinase, MMPs) to prevent degradation of collagen , and play a key role in the progression of liver fibrosis also promote collagen secretion by inhibiting apoptosis of HSC . Therefore, to reduce the expression of CTGF and TIMP-1 , can reduce the degree of liver fibrosis . This study aimed to construct the the rat CTGF or TIMP-1 gene targeting short hairpin RNA (short hairpin RNA, shRNA) expression vector , to provide a powerful tool to further explore the gene therapy of liver fibrosis . Methods: RNA interference target CTGF and TIMP-1 gene is the most effective pre- screened , that CTGF gene 1560 ~ 1580nt and TIMP -1 gene 412 ~~ 432nt RNA interference ( RNA interference , RNAi), sequence design principles each design containing short hairpin RNAi target sequences , annealing to form a double-stranded DNA , were cloned into the recombinant plasmid vector containing the purpose of the target gene fragment psiRNA-GFP construct by double digestion of the plasmid vector psiRNA - h7SKGFPzeo the -CTGF and psiRNA - of GFP- of TIMP- 1 , and the recombinant plasmids were double digested agarose gel electrophoresis and sequencing . Results: digestion confirmed to rat CTGF or TIMP-1 gene to target the expression of the shRNA target gene fragments were cloned into plasmid vector psiRNA-h7SKGFPzeo sequencing results show that the insertion sequence of the recombinant plasmid vector design target gene fragment is completely consistent . Conclusion : The successful construction of targeting rat CTGF and TIMP-1 of the most effective RNA interference target shRNA expression plasmid recombinant lay the experimental basis for further exploration of new ways of hepatic fibrosis gene therapy .

Related Dissertations

  1. Expression Dynamics of Pheromone Binding Proteins and Expression Influences by Mating and Knockdown of Cryl in Spodoptera Exigua,S433.4
  2. Molecular Cloning, Mrna Expression and Rnai of Nadph-Cytochrome P450 Reductase Gene in Helicoverpa Armigera (Hǘbner),S435.622.3
  3. Effects of BMPR-IB Gene Silencing by Small Interfering RNA on Apoptosis of Porcine Pollicular Granulosa Cells and Ecpression of BMP Pathy-Way-Ralted Genes,S828
  4. The DNA Cloning and RNA Interference of Some Functional Genes in Laodelphax Striatellus (Fallen),S435.112.3
  5. Silencing of SOCS1 Enhances Dc-mediated Anti-laryngocarcinoma Immunity,R739.65
  6. Knock-down of Cullin-5 Expression on Alteration of Morphology and Proliferation Ability in Glomerular Podocytes,R726.9
  7. The Roles and Mechanism of HIF-2α in Pathogenesis of Ischemic/Hypoxic Acute Kidney Injury,R692
  8. The Study of Both Magnetic Resonance Diffusion-weighted Imaging and Expression of VEGF Compared with Pathology in Dog Model of Liver Fibrosis,R575.2
  9. The Function Characterization of Salt-tolerant Correlative Gene TaSI1 and TaSC in Wheat,Q943
  10. Screening of Effective Fractions and Pharmacodynamical Study on Liver Protective Activity of Capparis Spinosa L.Fruits,R29
  11. The Effect of Inhibiting FoxO1 on the Glucose Consumption of Insulin Resistant Cells and the Mechanism,R587.1
  12. Bmi1 Regulates Self-Renewal and Tumorigenicity of Cancer Stem Cells in Non-Small Cell Lung Cancer,R734.2
  13. Correlation between HMGB1 and Cervical Cancer Metastasis,R737.33
  14. The Construction of Survivin shRNA Lentiviral Vector,R733.3
  15. Rabbit eyes trabeculectomy combined with amniotic membrane wrapped scleral flap surgical experimental study,R779.6
  16. Expression of Retinal CTGF and Intervention of Simvastatin in Rats with Diabetic Retinopathy,R774.1
  17. Effects of CTGF on Cardiomyocyte Hypertrophy of Rat and Study in Its Possible Signal Transduction Mechanism and Interfere Effect of Lacidipine,R965
  18. Chymasc Inhibitor in the Rat Model of DMN Induced Liver Fibrosis and Its Impact on the Expression of Angiotensin Ⅱ and Collagen Ⅰ,R965
  19. Molecular Cloning of ω-gliadin Genes and Study on the Expression of α-gliadin Gene Family by RNAi,S512.1
  20. Studies on the Construction of Two Chemical-inducible Expression Vectors and Transient Expression Analysis of VGCfE: Gus in Tobacco,S511
  21. RNA Interference and Overexpression of LXRα Gene on Fatty Acid Metabolism in the Mammary Gland Epithelial Cell of Dairy Goat,S827

CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Liver and gall bladder disease > Cirrhosis
© 2012 www.DissertationTopic.Net  Mobile