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Chinese Journal of Biotechnology ; (12): 1690-1696, 2009.
Article in Chinese | WPRIM | ID: wpr-296871

ABSTRACT

External Guide Sequences (EGSs) represents a novel nucleic acid based gene interference approach to modulate gene expression. They are oligonucleotides that consist of a sequence complementary to a target mRNA and recruit intracellular RNase P for specific degradation of the target RNA. DNA-based EGS1386 with a size of 12 nt was chemically synthesized to target the mRNA coding for the UL49 gene of human cytomegalovirus (HCMV). The DNA-based EGS1386 molecule efficiently directed human RNase P to cleave the target mRNA sequence in vitro. A reduction of more than 50% in the levels of UL49 expression was observed in human cells treated with the DNA-based EGS1386 targeted UL49 assayed by fluorescent quantization PCR and Western blotting. This results showed that the DNA-EGS1386 can effectively guide the RNase P cut the target mRNA. Therefore, DNA-EGS can develop into a new gene silencing technology and potential of the anti-viral reagents.


Subject(s)
Humans , Base Sequence , Cytomegalovirus , Genetics , Metabolism , Cytomegalovirus Infections , Virology , DNA, Viral , Genetics , Directed Molecular Evolution , Methods , Gene Expression Regulation, Viral , Nucleic Acid Conformation , Oligodeoxyribonucleotides , Genetics , Pharmacology , Chemistry , Pharmacology , RNA, Messenger , Genetics , Metabolism , Ribonuclease P , Genetics , Metabolism , Viral Structural Proteins , Genetics , Metabolism
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