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
在 En
| WPRIM
| ID: wpr-282072
Responsible library:
WPRO
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>
全文:
1
索引:
WPRIM
主要主题:
Pharmacology
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Physiology
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Plasmids
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Transfection
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Gene Expression
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Analysis of Variance
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Gene Silencing
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RNA, Small Interfering
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Cell Line, Tumor
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Green Fluorescent Proteins
研究类型:
Prognostic_studies
限制:
Animals
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Humans
语言:
En
期刊:
Neuroscience Bulletin
年:
2009
类型:
Article