Development of a novel trans-lentiviral vector that affords predictable safety.
Development of a novel trans-lentiviral vector that affords predictable safety.
复制标题
开发一种新型反式慢病毒载体，提供可预测的安全性。
DOI：
10.1006/mthe.2000.0095
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Kappes,JC
中科院分区：
文献类型：
--
作者：
Wu,X;Wakefield,JK;Liu,H;Xiao,H;Kralovics,R;Prchal,JT;Kappes,JC
Lentiviral vectors derived from human immunodeficiency virus type 1 (HIV-1) hold great promise for gene therapy. However, the possibility of generating replication-competent retrovirus (RCR) through genetic recombination raises concerns for safety. Here we describe a novel HIV-based packaging system (trans-lentiviral) that splits gag/gag-pol into two parts: One that expresses gag/gag-pro and another that expresses reverse transcriptase and integrase as fusion partners of viral protein R (Vpr). Using a sensitive assay developed to specifically detect recombinant lentiviral DNA mobilization, we demonstrated that the trans-lentiviral vector prevents the generation of recombinants that contain a functional gag-pol structure, while the lentiviral vector generatesenv-minus recombinant lentivirus that mobilizes recombinant genomes to other cells when pseudotyped with an exogenous envelope. Since an intact gag-pol structure is absolutely required for retroviral DNA mobilization and RCR, the trans-lentiviral vector design significantly reduces this risk. Moreover, it makes it possible to assess the risk of RCR and DNA mobilization using anin vitroassay that monitors trans-lentiviral vector stocks for the regeneration of the gag-pol structure. Therefore, the trans-lentiviral vector design will ensure the greatest predictable level of safety for the clinical application of retroviral vectors, including HIV-based vectors.
