Jones John Patrick, Kierlin Monique N, Coon Robert G, Perutka Jiri, Lambowitz Alan M, Sullenger Bruce A
Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA.
Mol Ther. 2005 May;11(5):687-94. doi: 10.1016/j.ymthe.2005.01.014.
Retroposable elements such as retroviral and lentiviral vectors have been employed for many gene therapy applications. Unfortunately, such gene transfer vectors integrate genes into many different DNA sequences and unintended integration of the vector near a growth-promoting gene can engender pathological consequences. For example, retroviral vector-mediated gene transfer induced leukemia in 2 of 11 children treated for severe combined immunodeficiency, raising significant safety issues for gene transfer strategies that cannot be targeted to specific sequences. Here, we examine the use of a mobile retroposable genetic element that can be targeted to introduce therapeutic sequences site specifically into mutant genes. The data demonstrate that the mobile group II intron from Lactococcus lactis can be targeted to insert into and repair mutant lacZ (approved gene symbol GLB1) and beta-globin (approved gene symbol HBB) genes with high efficiency and fidelity in model systems in bacteria. These results suggest that these mobile genetic elements represent a novel class of agents for performing targeted genetic repair.
逆转录病毒和慢病毒载体等可移动元件已被用于许多基因治疗应用中。不幸的是，这类基因转移载体将基因整合到许多不同的DNA序列中，并且载体在促进生长的基因附近意外整合可能会产生病理后果。例如，在接受严重联合免疫缺陷治疗的11名儿童中，有2名儿童因逆转录病毒载体介导的基因转移诱发了白血病，这给无法靶向特定序列的基因转移策略带来了重大安全问题。在此，我们研究了一种可移动的逆转录遗传元件的应用，该元件可被靶向用于将治疗性序列特异性地导入突变基因中。数据表明，来自乳酸乳球菌的可移动II组内含子可以在细菌模型系统中高效且准确地靶向插入并修复突变的lacZ（批准基因符号GLB1）和β-珠蛋白（批准基因符号HBB）基因。这些结果表明，这些可移动遗传元件代表了一类用于进行靶向基因修复的新型试剂。
