Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector
Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector
复制标题
DOI：
10.1073/pnas.93.21.11382
复制
发表时间：
1996-10-15
影响因子：
11.1
通讯作者：
Verma, IM
中科院分区：
文献类型：
--
作者：
Naldini, L;Blomer, U;Verma, IM
我们描述了一个安全的，复制缺陷型和有效的慢病毒载体，适合在体内基因传递的建设。发现载体的逆转录是限速步骤;因此，在递送之前促进载体颗粒内部的反应显著提高了基因转移的效率。在注射到成年大鼠的脑中后，在没有可检测到的病理的情况下获得转基因的持续长期表达。在载体注射部位周围区域的高比例神经元表达转导的β-半乳糖苷酶基因。这种模式在单次给予载体后几个月处死的动物中是不变的。转导通过载体基因组的整合而发生，因为它被整合到载体中的整合酶蛋白的催化位点中的单个氨基酸取代所消除。因此，慢病毒载体的临床上可接受的衍生物的开发可以使显著量的治疗性基因产物在多种体细胞组织中的持续递送成为可能。
We describe the construction of a safe, replication-defective and efficient lentiviral vector suitable for in vivo gene delivery. The reverse transcription of the vector was found to be a rate-limiting step; therefore, promoting the reaction inside the vector particles before delivery significantly enhanced the efficiency of gene transfer. After injection into the brain of adult rats, sustained long-term expression of the transgene was obtained in the absence of detectable pathology. A high proportion of the neurons in the areas surrounding the injection sites of the vector expressed the transduced beta-galactosidase gene. This pattern was invariant in animals sacrificed several months after a single administration of the vector. Transduction occurs by integration of the vector genome, as it was abolished by a single amino acid substitution in the catalytic site of the integrase protein incorporated in the vector. Development of clinically acceptable derivatives of the lentiviral vector may thus enable the sustained delivery of significant amounts of a therapeutic gene product in a wide variety of somatic tissues.