Department of Ophthalmology, University of Florida, Gainesville, FL, USA.
Mol Ther. 2011 Feb;19(2):293-301. doi: 10.1038/mt.2010.234. Epub 2010 Nov 2.
Vectors based on adeno-associated virus serotype 2 (AAV2) have been used extensively in many gene-delivery applications, including several successful clinical trials for one type of Leber congenital amaurosis in the retina. Many studies have focused on improving AAV2 transduction efficiency and cellular specificity by genetically engineering its capsid. We have previously shown that vectors-containing single-point mutations of capsid surface tyrosines in serotypes AAV2, AAV8, and AAV9 displayed significantly increased transduction efficiency in the retina compared with their wild-type counterparts. In the present study, we evaluated the transduction characteristics of AAV2 vectors containing combinations of multiple tyrosine to phenylalanine mutations in seven highly conserved surface-exposed capsid tyrosine residues following subretinal or intravitreal delivery in adult mice. The multiply mutated vectors exhibited different in vivo transduction properties, with some having a unique ability of transgene expression in all retinal layers. Such novel vectors may be useful in developing valuable new therapeutic strategies for the treatment of many genetic diseases.
基于腺相关病毒血清型 2（AAV2）的载体已广泛应用于许多基因传递应用中，包括在视网膜中针对一种莱伯先天性黑蒙症的几种成功的临床试验。许多研究集中在通过基因工程改造其衣壳来提高 AAV2 的转导效率和细胞特异性。我们之前已经表明，与野生型相比，包含血清型 AAV2、AAV8 和 AAV9 衣壳表面酪氨酸单点突变的载体在视网膜中显示出显著增加的转导效率。在本研究中，我们评估了包含七个高度保守的表面暴露衣壳酪氨酸残基中多个酪氨酸到苯丙氨酸突变的 AAV2 载体在成年小鼠中经视网膜下或玻璃体内给药后的转导特征。多突变载体表现出不同的体内转导特性，其中一些具有在所有视网膜层中转基因表达的独特能力。这种新型载体可能有助于开发许多遗传疾病治疗的有价值的新治疗策略。
