An Evolved Adeno-associated Viral Variant Enhances Gene Delivery and Gene Targeting in Neural Stem Cells
An Evolved Adeno-associated Viral Variant Enhances Gene Delivery and Gene Targeting in Neural Stem Cells
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DOI：
10.1038/mt.2010.287
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发表时间：
2011-04-01
影响因子：
12.4
通讯作者：
Schaffer, David V.
中科院分区：
文献类型：
--
作者：
Jang, Jae-Hyung;Koerber, James T.;Schaffer, David V.
干细胞的基因传递和基因靶向将成为基础科学和生物医学应用的一项高度可行的技术。腺相关病毒（AAV）载体已证明能够有效递送至大量细胞，但其在干细胞中的应用因转导效率低而受到限制。然而，由于其相当大的优势，工程化 AAV 递送系统以增强向干细胞的基因递送可能会对干细胞生物学和治疗产生影响。因此，使用多种不同的 AAV 衣壳文库（包括随机诱变、DNA 改组和随机肽插入变体），我们应用定向进化来创建“设计”AAV 载体，提高神经干细胞 (NSC) 的递送效率。这种进化产生了一种新的 AAV 变体，它在肝素结合位点内的三重刺突表面插入了选定的肽序列。重要的是，这种进化的 AAV 变体介导了向大鼠、小鼠和人类 NSC 的有效基因传递，以及成人 NSC 内的有效基因靶向，因此它在从基础干细胞生物学到临床转化的应用领域具有广阔的前景。
Gene delivery to, and gene targeting in, stem cells would be a highly enabling technology for basic science and biomedical application. Adeno-associated viral (AAV) vectors have demonstrated the capacity for efficient delivery to numerous cells, but their application to stem cells has been limited by low transduction efficiency. Due to their considerable advantages, however, engineering AAV delivery systems to enhance gene delivery to stem cells may have an impact in stem cell biology and therapy. Therefore, using several diverse AAV capsid libraries-including randomly mutagenized, DNA shuffled, and random peptide insertion variants-we applied directed evolution to create a "designer" AAV vector with enhanced delivery efficiency for neural stem cells (NSCs). A novel AAV variant, carrying an insertion of a selected peptide sequence on the surface of the threefold spike within the heparin-binding site, emerged from this evolution. Importantly, this evolved AAV variant mediated efficient gene delivery to rat, mouse, and human NSCs, as well as efficient gene targeting within adult NSCs, and it is thus promising for applications ranging from basic stem cell biology to clinical translation.