Retroviral pseudotransduction for targeted cell manipulation
Retroviral pseudotransduction for targeted cell manipulation
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DOI：
10.1016/j.molcel.2004.09.023
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发表时间：
2004-10-22
期刊：
影响因子：
16
通讯作者：
Baum, C
中科院分区：
文献类型：
--
作者：
Galla, M;Will, E;Baum, C
目前的研究解决了逆转录病毒载体是否可以被修改，以实现受体介导的，剂量控制的，和瞬时递送蛋白质或核酸到靶细胞。作为范例，我们产生了编码位点特异性重组酶Cre的基于小鼠白血病病毒的载体。载体在引物结合位点功能中被禁用，阻断病毒体mRNA的逆转录。虽然减少转基因插入超过1000倍，并取消组成型Cre表达的毒性作用，瞬时Cre交付仍然是高效的，受体限制，和不敏感的逆转录的药理学抑制。这种形式的Cre转移需要逆转录病毒包装信号，翻译单位的近端帽定位，以及生产细胞中的gag和env表达，揭示了逆转录病毒mRNA转移的潜在机制。因此，逆转录病毒递送的mRNA如果不被逆转录，则可以充当立即翻译模板。这种方法允许用核酸进行靶向和可逆的细胞操作的多种修饰。
The present study addressed whether retroviral vectors could be modified to achieve receptor-mediated, dose-controlled, and transient delivery of proteins or nucleic acids into targeted cells. As a paradigm, we generated mouse leukemia virus-based vectors encoding the site-specific recombinase Cre. The vectors were disabled in primer binding site function, blocking reverse transcription of the virion mRNA. While reducing transgene insertion more than 1000-fold and abolishing toxic effects of constitutive Cre expression, transient Cre delivery was still highly efficient, receptor restricted, and insensitive to pharmacologic inhibition of reverse transcription. This form of Cre transfer required the retroviral packaging signal, cap-proximal positioning of the translation unit, as well as gag and env expression in producer cells, revealing retroviral mRNA transfer as the underlying mechanism. Thus, retrovirally delivered mRNA may serve as an immediate translation template if not being reverse transcribed. This approach allows multiple modifications for targeted and reversible cell manipulation with nucleic acids.