Comparison of gene expression after intraperitoneal delivery of AAV2 or AAV5 in utero
Comparison of gene expression after intraperitoneal delivery of AAV2 or AAV5 in utero
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DOI：
10.1016/s1525-0016(03)00132-1
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发表时间：
2003-07-01
影响因子：
12.4
通讯作者：
Gaensler, KML
中科院分区：
文献类型：
--
作者：
Lipshutz, GS;Titre, D;Gaensler, KML
疾病的纠正可以通过将基因传递给干细胞和发育器官系统来实现。我们以前的研究表明，在子宫内注射腺相关病毒（AAV）血清型2在小鼠体内的终身表达。在本研究中，我们比较了在子宫内第15天胎儿中通过腹膜内注射与荧光素酶连接的AAV 2和AAV 5中使用延伸因子1 α（EF1 α）或CMV启动子的表达水平。另外的AAV构建体还含有土拨鼠肝炎病毒转录后调控元件（WPRE）。荧光素酶表达的水平和分布通过体内生物发光和发光测定来评估。所有小鼠表现出荧光素酶表达>15个月。在体内，来自AAV5的荧光素酶表达大于从AAV2产生的荧光素酶表达。含有CMV启动子的载体在检查的所有组织中产生了更高水平的基因表达，与EF1 α定向载体相比。WPRE使体外表达增加4倍，体内表达增加8倍。这些研究表明，通过修饰启动子和血清型，可以实现AAV指导的表达效率的增加。rAAV介导的子宫内基因递送的功效支持这些载体用于未来疗法的潜力。
Correction of diseases may be achieved by delivery of genes to stem cells and developing organ systems. Our previous studies demonstrated life-long expression after in utero injection of adeno-associated virus (AAV) serotype 2 in mice. In the present studies, we compared levels of expression using the elongation factor 1alpha (EF1alpha) or the CMV promoter in AAV2 and AAV5 linked to luciferase via intraperitoneal injection in day 15 fetuses in utero. An additional AAV construct also contained the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). The level and distribution of luciferase expression were assessed by in vivo bioluminescence and luminometric assays. All mice exhibited luciferase expression for >15 months. In vivo, luciferase expression from AAV5 was greater than that produced from AAV2. Vectors containing the CMV promoter produced higher levels of gene expression in all tissues examined compared to EF1alpha-directed vectors. The WPRE increased expression in vitro fourfold and in vivo eightfold. These studies demonstrate that by modifying the promoter and serotype, increases in the efficiency of AAV-directed expression may be achieved. The efficacy of rAAV-mediated gene delivery in utero supports the potential of these vectors for future therapies.