Abstract
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.
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Acknowledgments
This study was supported by the research grants to H.X. from the National Natural Science Foundation of China (Nos. 81272543 and 31070137).
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Li, X., Xia, R., Zhao, J. et al. A one-step ligation system for rapid generation of armed, conditionally-replicating adenoviruses. Biotechnol Lett 35, 1215–1221 (2013). https://doi.org/10.1007/s10529-013-1214-6
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DOI: https://doi.org/10.1007/s10529-013-1214-6