多能干细胞中的游离型转基因表达。

Episomal transgene expression in pluripotent stem cells.

作者信息

Lufino Michele M P, Popplestone Anna R, Cowley Sally A, Edser Pauline A H, James William S, Wade-Martins Richard

机构信息

Molecular Neurodegeneration and Gene Therapy Research Group, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.

出版信息

Methods Mol Biol. 2011;767:369-87. doi: 10.1007/978-1-61779-201-4_27.

DOI:10.1007/978-1-61779-201-4_27

PMID:21822889

Abstract

Herpes simplex type 1 (HSV-1) amplicon vectors possess a number of features that make them excellent vectors for the delivery of transgenes into stem cells. HSV-1 amplicon vectors are capable of efficiently transducing both dividing and nondividing cells and since the virus is quite large, 152 kb, it is of sufficient size to allow for incorporation of entire genomic DNA loci with native promoters. HSV-1 amplicon vectors can also be used to incorporate and deliver to cells a variety of sequences that allow extrachromosomal retention. These elements offer advantages over integrating vectors as they avoid transgene silencing and insertional mutagenesis. The construction of amplicon vectors carrying extrachromosomal retention elements, their packaging into HSV-1 viral particles, and the use of HSV-1 amplicons for stem cell transduction will be described.

摘要

单纯疱疹病毒1型（HSV-1）扩增子载体具有许多特性，使其成为将转基因导入干细胞的理想载体。HSV-1扩增子载体能够有效地转导分裂细胞和非分裂细胞，并且由于该病毒相当大，有152 kb，其大小足以允许整合带有天然启动子的整个基因组DNA位点。HSV-1扩增子载体还可用于整合并向细胞递送多种允许染色体外保留的序列。这些元件比整合载体具有优势，因为它们可避免转基因沉默和插入诱变。本文将描述携带染色体外保留元件的扩增子载体的构建、其包装到HSV-1病毒颗粒中以及使用HSV-1扩增子进行干细胞转导的方法。