Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an, 710062, People's Republic of China.
Biotechnol Lett. 2013 Aug;35(8):1215-21. doi: 10.1007/s10529-013-1214-6. Epub 2013 Apr 23.
Conditionally-replicating adenoviruses (CRAd) are becoming a promising strategy for cancer therapy. However, the generation of CRAd is complicated and time-consuming, especially for armed CRAd Ad5-D24. A one-step ligation system is now reported for rapid construction of an armed CRAd Ad5-D24 carrying two foreign transgenes. The system has the following features: (1) the CRAd backbone pCRAd5-D24-BS-Lacza contains two unique restriction sites between the fiber and E4 for easy introduction of a transgene expression cassette; and (2) a bi-directional promoter in pshuttle-BS-Dual allows for the expression of two independent transgenes. As proof of principle, Ad5-D24 CRAd carrying eGFP and mCherry was generated and examined. This novel ligation system will be a versatile and efficient tool in cancer viral gene therapy.
条件复制型腺病毒 (CRAd) 正成为癌症治疗的一种有前途的策略。然而，CRAd 的产生过程复杂且耗时，特别是对于武装型 CRAd Ad5-D24 而言。现在报道了一种一步连接系统，用于快速构建携带两种外源转基因的武装型 CRAd Ad5-D24。该系统具有以下特点：（1）CRAd 骨架 pCRAd5-D24-BS-Lacza 在纤维和 E4 之间包含两个独特的限制酶切位点，便于引入转基因表达盒；（2）pshuttle-BS-Dual 中的双向启动子允许两个独立的转基因表达。作为原理验证，生成并检测了携带 eGFP 和 mCherry 的 Ad5-D24 CRAd。这种新型连接系统将成为癌症病毒基因治疗的通用且高效的工具。
