Chai Sunghee, Wakefield Leslie, Norgard Mason, Li Bin, Enicks David, Marks Daniel L, Grompe Markus
Papé Family Pediatric Research Institute, Oregon Stem Cell Center, Portland, OR, USA.
Department of Pediatrics, Oregon Health & Science University, Portland, OR, USA.
Mol Ther Methods Clin Dev. 2023 May 17;29:504-512. doi: 10.1016/j.omtm.2023.05.013. eCollection 2023 Jun 8.
Significant progress has been made in developing recombinant adeno-associated virus (rAAV) for clinical gene therapy. While rAAV is a versatile gene delivery platform, its packaging limit of 4.7 kb limits the diseases it can target. Here, we report two unusually small promoters that enable the expression of larger transgenes than standard promoters. These micro-promoters are only 84 (MP-84) and 135 bp (MP-135) in size but have activity in most cells and tissues comparable to the CAG promoter, the strongest ubiquitous promoter to date. MP-84- and MP-135-based rAAV constructs displayed robust activity in cultured cells from the three different germ-layer lineages. In addition, reporter gene expression was documented in human primary hepatocytes and pancreatic islets and in multiple mouse tissues , including brain and skeletal muscle. MP-84 and MP-135 will enable the therapeutic expression of transgenes currently too large for rAAV vectors.
在开发用于临床基因治疗的重组腺相关病毒（rAAV）方面已取得重大进展。虽然rAAV是一种通用的基因递送平台，但其4.7 kb的包装限制限制了它能够靶向治疗的疾病。在此，我们报告了两个异常小的启动子，它们能够表达比标准启动子更大的转基因。这些微型启动子大小仅为84（MP-84）和135 bp（MP-135），但在大多数细胞和组织中的活性与CAG启动子相当，CAG启动子是迄今为止最强的普遍存在的启动子。基于MP-84和MP-135的rAAV构建体在来自三种不同胚层谱系的培养细胞中显示出强大的活性。此外，在人原代肝细胞和胰岛以及包括脑和骨骼肌在内的多种小鼠组织中记录到了报告基因的表达。MP-84和MP-135将能够对目前对于rAAV载体而言过大的转基因进行治疗性表达。
