Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences
Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences
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DOI：
10.1038/76095
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发表时间：
2000-06-01
期刊：
影响因子：
30.8
通讯作者：
Naldini, L
中科院分区：
文献类型：
--
作者：
Follenzi, A;Ailles, LE;Naldini, L
基于慢病毒的基因转移载体的特征在于它们能够使非分裂细胞增殖（1，2）。HIV-1蛋白基质、Vpr和整合酶与非分裂细胞中病毒基因组的核输入有关（3-5）。在这里，我们表明，一个序列内的城邦也需要在顺式。它包含以前与靶细胞中逆转录进程相关的结构元件（6-9）。我们在后一代慢病毒载体中顺式恢复了这些元件（10，11）。新的载体转导效率高得多的几种类型的人类原代细胞，当生长和生长停滞，包括造血干细胞检测NOD/SCID小鼠的长期再增殖。在体内施用到SCID小鼠中时，载体诱导比未修饰的载体更高的人凝血因子IX（F.IX）的血浆水平。我们的研究结果表明，核易位的基因组是一个限速步骤慢病毒感染的分裂和非分裂细胞，它取决于蛋白质和核酸序列的决定因素。在基于慢病毒的载体中完全拯救这一步骤提高了基因治疗应用的性能。
Gene-transfer vectors based on lentiviruses are distinguished by their ability to transduce non-dividing cells(1,2). The HIV-1 proteins Matrix, Vpr and Integrase have been implicated in the nuclear import of the viral genome in non-dividing cells(3-5). Here we show that a sequence within polis also required in cis. It contains structural elements previously associated with the progress of reverse transcription in target cells(6-9). We restored these elements in cis within late-generation lentiviral vectors(10,11). The new vector transduced to a much higher efficiency several types of human primary cells, when both growing and growth-arrested, including haematopoietic stem cells assayed by longterm repopulation of NOD/SCID mice. On in vivo administration into SCID mice, the vector induced higher plasma levels of human clotting factor IX (F.IX) than non-modified vector. Our results indicate that nuclear translocation of the genome is a rate-limiting step in lentiviral infection of both dividing and non-dividing cells, and that it depends on protein and nucleic acid sequence determinants. Full rescue of this step in lentivirus-based vectors improves performance for gene-therapy applications.