Construction of VEGF gene-targeted small interfering RNA mediated by CMV promoter and its silencing effect / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 181-186, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-259220
ABSTRACT
<p><b>OBJECTIVE</b>To construct VEGF gene-targeted small interfering RNA (siRNA) and its expression vector driven by CMV promoter and to investigate its interference effect.</p><p><b>METHODS</b>The VEGF gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strands were synthesized. After annealing, two-strand oligonucleotide was inserted into pDC311-SV40-RC vector, which was then identified by PCR and sequenced. Then human U-2 OS cell line was transfected with the vector using lipofectamine method. Finally, ELISA was performed to evaluate the expression of VEGF protein.</p><p><b>RESULTS</b>PCR-identification of positive clone and sequencing confirmed the vector containing the target siRNA. ELISA showed that compared with the control group, the expression levels of VEGF protein in transfected U-2 OS cells were decreased significantly (P<0.05).</p><p><b>CONCLUSION</b>VEGF gene-targeted siRNA and its vector mediated by CMV promoter were successfully constructed, which can reduce the VEGF protein expression after transfecting.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Plasmids
/
Bone Neoplasms
/
RNA, Messenger
/
Molecular Sequence Data
/
Base Sequence
/
Transfection
/
Osteosarcoma
/
Adenoviruses, Human
/
Promoter Regions, Genetic
Type of study:
Prognostic study
Limits:
Humans
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2010
Type:
Article
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