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Experimental Study of the Sense Oligonucleotide on the Inhibition of Collagen Genes’ Promoter

Author: GaoYun
Tutor: YangSongLin
School: Shanghai Jiaotong University
Course: Surgical
Keywords: Sense oligonucleotide Transcription factor Gene Collagen Keloid Fibroblast
CLC: R622
Type: Master's thesis
Year: 2007
Downloads: 47
Quote: 0
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Abstract


Objective: To establish a sense oligonucleotide fragments of exogenous inhibition of collagen promoter related gene expression methods of observation , to explore the application of exogenous sense oligonucleotide inhibits the possibility of human scar collagen gene transcription . Materials and methods : human skin keloid fibroblasts primary culture ; construct containing human α1 (I) collagen gene promoter -2 483 --- 42bp sequence PGL - 3 reporter gene recombinant ; synthetic transcription factor Sp1, Ap1 specific recognition of double-stranded oligonucleotide sequence ; liposomes PGL - 3 reporter gene recombinant transfection keloid fibroblasts as a control , the experimental group transfected Sp- 1 , Ap- sense oligonucleotide acid fragment luciferase Assay system to detect the expression of this recombinant luciferase . Results: of PGL3-2. 5k and the predetermined value Ap1 , Sp1 sense oligo nucleotide double stranded can be transfected scar fibroblasts ; PGL3- 2.5k recombinant promoter sequence and Sp1 , the predetermined value Ap1 specifically recognize double-stranded oligonucleotide nucleoside recombinant acid can Sp1, Ap1 specific binding occurs . Experimental group ( of PGL3-2. 5k and the predetermined value Ap1 , Sp1 justice double-stranded oligonucleotide transfection ) and control group ( only transfection PGL3-2. 5k) luciferase expression compared , the difference was significant (P <0.01). CONCLUSION : The recombinant PGL3-2. 5k can be used for the promoter activity in keloid fibroblasts ; PGL3 report gene recombinant high sensitivity is very suitable for the α1 (I) collagen gene promoter activity in research ; collagen promoter activity decreased after the the sense oligonucleotide introduction of scar fibroblasts , experimental results show that the application of the sense oligonucleotide technology may be through the inhibition of collagen gene transcription activation , pathological scar dysplasia control .

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