Lee Seong-Wook, Jeong Jin-Sook
Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Yongin, South Korea.
Methods Mol Biol. 2014;1103:83-95. doi: 10.1007/978-1-62703-730-3_7.
One of the major concerns with regard to successful cancer gene therapy is to enhance both efficacy and safety. Gene targeting may represent an attractive tool to combat cancer cells without damage to normal cells. Here, we introduce a tumor-targeting approach with the Tetrahymena group I intron-based trans-splicing ribozyme, which cleaves target RNA and trans-ligate an exon tagged at the end of the ribozyme onto the downstream U nucleotide of the cleaved target RNA. We develop a specific trans-splicing ribozyme that can target and reprogram human cytoskeleton-associate protein 2 (hCKAP2)-encoding RNA to trigger therapeutic transgene herpes simplex virus thymidine kinase (HSVtk) selectively in cancer cells that express the RNA. Adenoviral vectors encoding the hCKAP2-specific trans-splicing ribozyme are constructed for in vivo delivery into either subcutaneous tumor xenograft or orthotopically multifocal hepatocarcinoma. We present analyses of the efficacy of the recombinant adenoviral vectors in terms of cancer retardation, target RNA and cell specificity, and in vivo toxicity.
成功进行癌症基因治疗的主要关注点之一是提高疗效和安全性。基因靶向可能是一种在不损害正常细胞的情况下对抗癌细胞的有吸引力的工具。在此，我们介绍一种基于四膜虫I组内含子的反式剪接核酶的肿瘤靶向方法，该核酶可切割靶RNA，并将标记在核酶末端的外显子反式连接到切割后的靶RNA下游的U核苷酸上。我们开发了一种特异性反式剪接核酶，它可以靶向并重新编程编码人细胞骨架相关蛋白2（hCKAP2）的RNA，从而在表达该RNA的癌细胞中选择性地触发治疗性转基因单纯疱疹病毒胸苷激酶（HSVtk）。构建了编码hCKAP2特异性反式剪接核酶的腺病毒载体，用于体内递送至皮下肿瘤异种移植或原位多灶性肝癌中。我们对重组腺病毒载体在癌症抑制、靶RNA和细胞特异性以及体内毒性方面的疗效进行了分析。
