title: A one-step ligation system for rapid generation of armed, conditionally-replicating adenoviruses.
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.
authors.affiliation: 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.
authors.forename: Xing
authors.initials: X
authors.lastname: Li
authors.forename: Ruiting
authors.initials: R
authors.lastname: Xia
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Dongyang
authors.initials: D
authors.lastname: Wang
authors.forename: Qinwen
authors.initials: Q
authors.lastname: Mao
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D009369:Neoplasms / Q000628:therapy