Development of an adenovirus vector lacking the expression of virus-associated RNAs
Development of an adenovirus vector lacking the expression of virus-associated RNAs
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DOI：
10.1016/j.jconrel.2011.06.020
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发表时间：
2011-09-25
影响因子：
10.8
通讯作者：
Mizuguchi, Hiroyuki
中科院分区：
文献类型：
--
作者：
Machitani, Mitsuhiro;Katayama, Kazufumi;Mizuguchi, Hiroyuki
使用腺病毒（Ad）载体的主要限制是先天免疫应答，其导致炎性细胞因子产生和组织损伤。为了克服这一限制，有必要开发更安全的Ad载体，其不太可能诱导先天免疫。Ad基因组编码两种非编码小RNA，病毒相关（VA）-RNA I和VA-RNA II，其由RNA聚合酶III转录并促进Ad复制。最近，我们报道了VA-RNA在用常规的第一代（E1缺失的）Ad载体（FG-Ad）转导的细胞中产生，并通过细胞内核酸传感器触发先天免疫应答。在本研究中，我们已经开发了一个VA-RNA-deleted Ad（Ad Delta VR）载体，其中VA-RNA表达的转录控制元件被删除。尽管常规HEK 293细胞不支持Ad Delta VR载体的增殖，但诱导表达VA-RNA I的HEK 293转化体（VR 293细胞）在适当诱导VA-RNA I表达的情况下允许Ad Delta VR载体的增殖。Ad Delta VR载体在培养的细胞中显示出与常规FG-Ad载体相当的高转导效率。Ad Delta VR载体可以是FG-Ad载体的更安全的替代物。(C)2011 Elsevier B. V.保留所有权利。
A major limitation of the use of adenovirus (Ad) vectors is the innate immune response, which causes inflammatory cytokine production and tissue damages. To overcome this limitation, it is necessary to develop safer Ad vectors that are less likely to induce innate immunity. The Ad genome encodes two non-coding small RNAs, virus-associated (VA)-RNA I and VA-RNA II, which are transcribed by RNA polymerase III and promote Ad replication. Recently, we reported that VA-RNAs are produced in the cells transduced with a conventional first-generation (El-deleted) Ad vector (FG-Ad) and trigger innate immune responses through intracellular nucleic acid sensors. In the present study, we have developed a VA-RNA-deleted Ad (Ad Delta VR) vector, in which the transcriptional control elements of the VA-RNA-expression were deleted. Although conventional HEK293 cells did not support the propagation of the Ad Delta VR vectors, HEK293 transformants inducibly expressing VA-RNA I (VR293 cells) with appropriate induction of VA-RNA I expression allowed the propagation of the Ad Delta VR vector. The Ad Delta VR vector showed high transduction efficiency comparable to that of the conventional FG-Ad vector in the cultured cells. The Ad Delta VR vector may be a safer alternative to the FG-Ad vector. (C) 2011 Elsevier B.V. All rights reserved.