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Abstract：
Dengue is a notorious viral infection, which affects a large segment of world populations in absence of vaccines and anti-viral treatment. The current study evaluates role of effective siRNA in dengue virus replication. Eight siRNA were synthesized against five different genes (Capsid, CprM, NS1, NS3 and NS5) of all serotypes of dengue virus. All serotype of DV were transfected with all synthesized siRNA in vitro, using BHK-21 cell lines. Culture fluid from test and control was tested by Real time PCR for CT value comparison in siRNA treated cell line (test) and untreated cell line (controls). Percent knockdown (%KD) was calculated by ∆∆CT methods to know the difference in test and control CT value. It was found that siRNA targeted against capsid gene worked best and showed inhibition of all four DV serotypes. DV-1, DV-2, DV-3 and DV-4 showed 93.8%, 99.3%, 87.5% and 93.8% knock down (%KD) respectively by siRNA targeted against capsid gene. Additionally, Si2 (target CprM gene 60-899) and Si 6 (target NS1 gene 3007-3025) were also showing inhibition of replication. Most serotypes of DV (with few exceptions) were not inhibited by siRNA targeted against NS-1, NS-3, and NS-5 genes. Animal studies using siRNAs are warranted to establish their therapeutic role.
摘要：
登革热是一种臭名昭著的病毒感染，在没有疫苗和抗病毒治疗的情况下，这影响了很大一部分世界人口。本研究评估了有效siRNA在登革病毒复制中的作用。针对五个不同的基因合成了八个siRNA（Capsid，Cprm,登革热病毒所有血清型的NS1，NS3和NS5）。用所有合成的siRNA体外转染所有血清型的DV,使用BHK-21细胞系。通过实时PCR测试来自测试和对照的培养液在siRNA处理的细胞系(测试)和未处理的细胞系(对照)中的CT值比较。通过ΔΔCT方法计算击倒百分比(%KD)以了解测试和对照CT值的差异。发现靶向衣壳基因的siRNA效果最好并且显示对所有四种DV血清型的抑制。DV-1，DV-2，DV-3和DV-4显示为93.8％，99.3%,通过靶向衣壳基因的siRNA分别降低87.5%和93.8%(%KD)。此外,Si2(靶CprM基因60-899)和Si6(靶NS1基因3007-3025)也显示复制抑制。大多数DV血清型(除了少数例外)不被靶向NS-1、NS-3和NS-5基因的siRNA抑制。使用siRNA的动物研究有必要确立其治疗作用。
