SiRNA structural optimization

人乳头瘤病毒(HPV)是导致肛门、头颈部肿瘤和恶性肿瘤的主要原因。 不幸的是，目前还没有针对HPV感染或HPV引起的恶性肿瘤的特效抗病毒疗法。 这份U19提案是Alexander在题为siRNA杀微生物剂的财团提案中作为项目提交的。这样做的目的是 研究联盟将开发天然的和化学修饰的siRNA作为阻断的有效手段 艾滋病毒、单纯疱疹病毒2型和人乳头瘤病毒的传播。在Alexander的项目中，我们假设化学修饰的单链 以硼磷为骨架的短干扰RNA(SiRNAs)在效力和持久性方面优于天然siRNA 基因沉默，以及抗HPV E6/E7单链硼磷酸盐骨架siRNAs(ssBP-siRNAs)可以 用于有效阻断HPV病毒复制和HPV转化细胞的增殖。通过将化学物质 为了增加siRNA沉默的效力和持续时间，我们将创建ssBP-siRNA， 设计用于局部给药，阻止HPV感染细胞中的Episome复制并诱导不可逆转的生长 在HPV16转化的细胞中发生停滞。我们鉴定的抗HPV16单链BP-siRNAs将在项目3中使用新颖的 人乳头瘤病毒感染的细胞培养和转基因小鼠模型。 项目3还将向Ramratnam和Herold的项目提供优化的ssBP-siRNA，以便将ssBP-siRNA与 用于靶向表达猕猴CCR5、鼠粘着斑激酶和鼠Nectin-1的常规siRNAs。 为了立即优化ssBP-sirna对阴道粘膜的输送，我们将为您提供现有的高活性 SSBP-siRNAs到核心B(配方)。配制的ssBP-siRNAs的阴道粘膜摄取和活性将是 在项目3中进行了定量评估。 在这项研究结束时，我们将开发出抗HPV16E6/E7ssBP-siRNAs作为有效的局部治疗药物 用于预防人乳头瘤病毒感染和人乳头瘤病毒诱发恶性肿瘤的杀微生物剂。我们还将创造出高效的 抗其他联盟靶标的ssBP-siRNAs，并将这些ssBP-siRNAs转发给 病毒感染的实验室模型。

Human papillomaviruses (HPVs) are a major cause of anogenital and head and neck neoplasms and malignancies. Unfortunately, no specific antiviral therapies for HPV infection or HPV-induced malignancies have been identified. This U19 proposal is submitted as project by Alexander in a consortium proposal entitled siRNA Microbicides. The goal of this research consortium is to develop native and chemically modified siRNAs as effective means for blocking transmission of HIV, HSV2 and HPV. In project by Alexander, we hypothesize that chemically modified single-stranded boranophosphate-backbone short interfering RNAs (siRNAs) are superior to native siRNAs for potent and durable gene silencing, and that anti-HPV E6/E7 single-stranded boranophosphate-backbone siRNAs (ssBP-siRNAs) can be used to effectively block HPV viral replication and proliferation of HPV-transformed cells. By combining chemical modifications shown individually to increase siRNA silencing potency and duration, we will create ssBP-siRNAs, designed for topical delivery, that block episome replication in HPV-infected cells and induce irreversible growth arrest in HPV16-transformed cells. Anti-HPV16 ssBP-siRNAs identified by us will be tested in project 3 using novel cell culture and transgenic mouse models of HPV infection. Project 3 will also supply optimized ssBP-siRNAs to projects by Ramratnam and Herold to allow comparison of ssBP-siRNAs with conventional siRNAs for targeting expression of macaque CCR5, murine focal adhesion kinase, and murine nectin-1. To allow immediate work optimizing ssBP-siRNA delivery to vaginal mucosa, we will supply existing highly active ssBP-siRNAs to Core B (Formulations). The vaginal mucosal uptake and activity of formulated ssBP-siRNAs will be assessed quantitatively in project 3. At the conclusion of this study, we will have developed anti-HPV16 E6/E7 ssBP-siRNAs as effective topical microbicides for prevention of HPV infection and HPV-induced malignancies. We will also have created high potency ssBP-siRNAs active against other consortium targets, and forwarded these ssBP-siRNAs for microbicidal testing in laboratory models of viral infection.