The construction of siRNA plasmid targeting mouse HIF-1alpha and in vitro study of its inhibition effect / 神经科学通报·英文版
Neuroscience Bulletin
;
(6): 122-130, 2009.
Article
in English
| WPRIM
| ID: wpr-282072
ABSTRACT
<p><b>OBJECTIVE</b>To construct effective RNA-interference plasmids targeting mouse HIF-1alpha gene and testify their effects and specificities in interfering HIF-1alpha expression.</p><p><b>METHODS</b>Three RNA-interference plasmids targeting mouse HIF-1alpha gene, pBS/U6/HIF-1alpha-siRNAI~III, were constructed and identified using double digestion method in the present study. RT-PCR, immunostaining and western blotting were employed to detect the expression alterations of HIF-1alpha in 293T cells following transfections of the three plasmids, respectively. The interference effect of pBS/U6/HIF1alphai-II in SH-SY5Y cell line was further investigated.</p><p><b>RESULTS</b>All the three RNA-interference plasmids, especially pBS/U6/HIF1alphai-II, showed significant inhibition in HIF-1alpha expression in 293T cell line. pBS/U6/HIF1alphai-II could also inhibit HIF-1alpha expression in SH-SY5Y cell line, in a dose-dependent way.</p><p><b>CONCLUSION</b>Plasmid pBS/U6/HIF1alphai-II constructed in our study can effectively and specifically inhibit HIF-1alpha expression, and its role in neural tube development and dysfunction will be further investigated. Construct of pBS/U6/HIF1alphai-II plasmid will provide a useful tool to study the role of HIF-1 pathway in embryogenesis, oncogenesis and ischemia development.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Physiology
/
Plasmids
/
Transfection
/
Gene Expression
/
Analysis of Variance
/
Gene Silencing
/
RNA, Small Interfering
/
Cell Line, Tumor
/
Green Fluorescent Proteins
Type of study:
Prognostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
Neuroscience Bulletin
Year:
2009
Type:
Article
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