title: A single-step mixing cloning method for assembly of lentiviral short hairpin RNA expression vectors for gene silencing.
abstract: Lentiviral expression vectors encoding short hairpin RNA (shRNA) are widely used for RNAi-based gene silencing in mammalian cells. However, current methods for the construction of shRNA expression vectors require multiple steps, which are expensive, time-consuming, and error-prone. Here, we developed a single-step mixing cloning method for the generation of lentiviral shRNA expression vectors. With this method, a pair of short oligonucleotides (∼50 nt) is required and a lentiviral shRNA vector can be constructed with only one step. This method has been used to construct 30 lentiviral shRNA expression vectors successfully.
authors.affiliation: Institute of Biochemistry and Molecular Biology, Hubei University, Wuhan 430062, Hubei Province, People's Republic of China.
authors.forename: Xing
authors.initials: X
authors.lastname: Zhong
authors.forename: Chao
authors.initials: C
authors.lastname: Zhai
authors.forename: Dengxiang
authors.initials: D
authors.lastname: Yang
authors.forename: Sijing
authors.initials: S
authors.lastname: Jiang
authors.forename: Zhezhe
authors.initials: Z
authors.lastname: Li
authors.forename: Xiaolan
authors.initials: X
authors.lastname: Yu
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Yapin
authors.initials: Y
authors.lastname: Wang
authors.forename: Wanping
authors.initials: W
authors.lastname: Chen
authors.forename: Lixin
authors.initials: L
authors.lastname: Ma
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D012097:Repressor Proteins / Q000172:deficiency / Q000235:genetics
chemical_list: D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C509761:SPOP protein, human