Targeted CNS Delivery Using Human MiniPromoters and Demonstrated Compatibility with Adeno-Associated Viral Vectors.
Targeted CNS Delivery Using Human MiniPromoters and Demonstrated Compatibility with Adeno-Associated Viral Vectors.
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DOI：
10.1038/mtm.2013.5
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发表时间：
2014-01-08
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影响因子：
--
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中科院分区：
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对于人类基因治疗来说，关键是有可用的小启动子工具，以高度特异和可重复性的方式驱动基因表达。我们利用计算生物学和系统发育保守的方法开发了人类DNA微型启动子(MiniPs)，从而解决了这一挑战。在小鼠身上检测X染色体上HPRT基因座的单拷贝敲入，并评估LacZ报告基因在中枢神经系统(CNS)和非中枢神经系统组织中的表达。在小鼠脑内鉴定出18个新的小分子蛋白驱动表达，2个小分子蛋白驱动泛神经元表达，17个小鼠眼睛蛋白表达。在这一组中代表了治疗感兴趣的关键区域：大脑皮层、胚胎下丘脑、脊髓、视网膜的双极细胞和神经节细胞，以及骨骼肌。我们还证明了三个视网膜神经节细胞MIFP在通过玻璃体内的腺相关病毒载体输送时表现出相似的细胞类型特异性。我们得出的结论是，我们的方法和特征已经导致了预期的表达特征，这些特征是微型启动子固有的，而不是由拷贝数效应或基因组位置决定的，并且导致了在腺相关病毒中容易成功的构建。这些小分子药物立即适用于人类基因治疗的临床前研究，并可公开用于促进基础和临床研究以及人类基因治疗。
Critical for human gene therapy is the availability of small promoters tools to drive gene expression in a highly specific and reproducible manner. We tackled this challenge by developing human DNA MiniPromoters (MiniPs) using computational biology and phylogenetic conservation. MiniPs were tested in mouse as single-copy knock-ins at the Hprt locus on the X chromosome and evaluated for lacZ reporter expression in central nervous system (CNS) and non–CNS tissue. Eighteen novel MiniPs driving expression in mouse brain were identified, 2 MiniPs for driving pan-neuronal expression and 17 MiniPs for the mouse eye. Key areas of therapeutic interest were represented in this set: the cerebral cortex, embryonic hypothalamus, spinal cord, bipolar and ganglion cells of the retina, and skeletal muscle. We also demonstrated that three retinal ganglion cell MiniPs exhibit similar cell type specificity when delivered via adeno-associated virus vectors intravitreally. We conclude that our methodology and characterization has resulted in desirable expression characteristics that are intrinsic to the MiniPromoter, not dictated by copy-number effects or genomic location, and results in constructs predisposed to success in adeno-associated virus. These MiniPs are immediately applicable for preclinical studies toward gene therapy in humans and are publicly available to facilitate basic and clinical research, and human gene therapy.
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影响因子：
56.9
作者：
Carninci, P;Kasukawa, T;Hayashizaki, Y
通讯作者：
Hayashizaki, Y
影响因子：
5.3
作者：
Forni, Paolo E.;Scuoppo, Claudio;Ponzetto, Carola
通讯作者：
Ponzetto, Carola
影响因子：
4.2
作者：
Cordier, L;Gao, GP;Sweeney, HL
通讯作者：
Sweeney, HL
DOI：
10.1073/pnas.0409103102
发表时间：
2005-01-25
影响因子：
11.1
作者：
Li, SJ;Czubryt, MP;Olson, EN
通讯作者：
Olson, EN
影响因子：
7
作者：
Kheradpour, Pouya;Ernst, Jason;Kellis, Manolis
通讯作者：
Kellis, Manolis