使用睡美人转座子系统的肝脏定向基因治疗。

Liver-directed gene therapy using the sleeping beauty transposon system.

作者信息

Belur Lalitha R, McIvor R Scott, Wilber Andrew

机构信息

Arnold and Mabel Beckman Center for Transposon Research, Gene Therapy Program, Institute of Human Genetics, and Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA.

出版信息

Methods Mol Biol. 2008;434:267-76. doi: 10.1007/978-1-60327-248-3_16.

DOI:10.1007/978-1-60327-248-3_16

PMID:18470650

Abstract

Sleeping Beauty (SB) is a transposon system genetically reconstructed from teleost fish that mediates chromosomal integration of DNA sequences by a cut-and-paste mechanism. SB has been shown to mediate transposition in a variety of cells and tissues, has been used for the generation of transgenic animals and has been tested as a vector for gene therapy in several animal models of human disease. Here, we describe methods that we have developed for testing SB-mediated transposition, first in cultured mammalian cells, and then in vivo using a combination of rapid, high-volume tail vein injection for DNA delivery to the liver along with in vivo bioluminescence imaging to monitor sustained luciferase gene expression in individual animals.

摘要

睡美人（SB）是一种从硬骨鱼中基因重建的转座子系统，它通过剪切粘贴机制介导DNA序列的染色体整合。已证明SB可在多种细胞和组织中介导转座，已用于生成转基因动物，并已在几种人类疾病动物模型中作为基因治疗载体进行了测试。在这里，我们描述了我们开发的用于测试SB介导的转座的方法，首先是在培养的哺乳动物细胞中，然后是在体内，使用快速、大容量尾静脉注射将DNA输送到肝脏，并结合体内生物发光成像来监测个体动物中荧光素酶基因的持续表达。