Iranian Biomedical Journal
مجله بیومدیکال ایران
IBJ
Basic Sciences
http://ibj.pasteur.ac.ir
1
admin
1028-852X
2008-823X
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10.61186/ibj
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8888
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en
jalali
1387
10
1
gregorian
2009
1
1
13
1
online
1
fulltext
en
افزایش اثر تداخل RNA در سلول های P19 ECبه وسیله آنزیم RNA پلی مراز وابسته به RNA
Enhancement of RNA Interference Effect in P19 EC Cells by an RNA-dependent RNA Polymerase
Related Fields
Related Fields
مقاله کامل
Full Length/Original Article
Background: RNA interference (RNAi) is a phenomenon uses double-stranded RNA (dsRNA) to specifically inhibit gene expression. The non-specific silencing caused by interferon response to dsRNA in mammalian cells limits the potential of utilizing RNAi to study gene function. Duplexes of 21-nucleotide short interfering dsRNA (siRNA) inhibit gene expression by RNAi. In some organisms, siRNA can also function as a primer converting mRNA into dsRNA that are further cleaved to produce more siRNA. This activity involves the enzyme RNA-dependent RNA polymerase (RdRP). There are no known RdRP involved in RNAi in mammals. By using an RdRP from Caenorhabditis elegance named ego-1, investigators intend to enhance RNAi effect in mammalian cells. The aims of this project were: 1) to investigate the efficiency of siRNA to enhanced green fluorescent protein (eGFP) gene silencing and 2) to enhance the RNAi effect. Methods: We used a vector-based siRNA to target eGFP. Also we used a vector expressing ego-1 to test for a possible amplification effect of RNAi. The expression of eGFP in the cells was detected by using fluorescent microscopy, flowcytometry and Western-blotting. Results: Transfection of the plasmid into P19 cells significantly decreased eGFP fluorescence. In addition, eGFP protein was reduced. Preliminary data suggested that the presence of ego-1 enhanced the RNAi effect. Conclusion: The results indicated that use of hairpin siRNA expression vectors for RNAi is a promising method to inhibition of gene expression in mammalian cells. Also, introducing RdRP enzyme to mammalian cells might amplify the RNAi effect in the cells.
Gene silencing, RNA interference (RNAi), RNA-dependent RNA polymerase (RdRP), Double-stranded RNA (dsRNA)
19
25
http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-16&slc_lang=en&sid=1
Fariba
Esmaeili
فریبا
اسماعیلی
esmail_far@yahoo.com
100319475328460056922
100319475328460056922
Yes