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Production of pantropic retrovirus by ecotropic packaging cell line

机译：通过嗜性包装细胞系生产全向性逆转录病毒

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The range of infectivity (tropism) of the packaged retrovirus is generally determined by envprotein expressed in packaging cell line. Therefore, conversion of the env component of existing packaging cell line to another one can alter the tropism of packaged retrovirus. To achieve this, the VSV-G vector encoding the VSV-G env glycoprotein was cotransfected with the retroviral GFP expression vector into BOSC23 ecotropic packaging cell line which stably express the MMLV env protein. Unfortunately, the produced virus could only infect few human 293F cells, indicating the VSV-G can not totally replace the MMLV env as an env component of the packaged virus in BOSC23 cells. Therefore, RNAi technology was used to silence the MMLV env gene expression. The high-titer pantropic retrovirus were obtained by cotransfection of BOSC23 cells with MMLV env silencing vector, VSV-G vector, and retroviral GFP expression vector. When the pantropic retrovirus was applied to gene transfer of human umbilical cord blood CD34+ cells, the enriched CD34+ cells were successfully transduced. These results suggest that a method by which different tropic (pantropic) retrovirus can be packaged using an existing, tropism-fixed (ecotropic) packaging cell line has been successfully developed. This method is especially useful for researchers in developing countries who have one packaging cell line with tropism not compatible with the target cells and can not easily obtain the comercial packaging cell lines because there might not be enough suppliers to provide the commercial products of packaging cell lines in developing countries.
机译：包装的逆转录病毒的感染性（向性）范围通常由包装细胞系中表达的环境蛋白决定。因此，将现有包装细胞系的env组分转化为另一种可以改变包装的逆转录病毒的嗜性。为此，将编码VSV-G env糖蛋白的VSV-G载体与逆转录病毒GFP表达载体共转染到稳定表达MMLV env蛋白的BOSC23嗜温包装细胞系中。不幸的是，产生的病毒只能感染少数人293F细胞，这表明VSV-G不能完全取代MMLV env作为BOSC23细胞中包装病毒的env成分。因此，RNAi技术被用于沉默MMLV env基因表达。高滴度全嗜逆转录病毒是通过将BOSC23细胞与MMLV env沉默载体，VSV-G载体和逆转录病毒GFP表达载体共转染而获得的。当将全向逆转录病毒用于人脐带血CD34 +细胞的基因转移时，成功地转导了富集的CD34 +细胞。这些结果表明，已经成功开发了一种方法，该方法可以使用现有的，向性固定的（共亲性）包装细胞系包装不同的热带（泛性）逆转录病毒。这种方法对发展中国家的研究人员特别有用，因为他们的包装细胞系具有与目标细胞不兼容的向性，并且由于可能没有足够的供应商提供包装细胞系的商业产品而无法轻易获得商业包装细胞系。在发展中国家。

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