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Updated: May 13, 2026
Lentivirus Production
Published on: October 2, 2009
A single-step mixing cloning method for assembly of lentiviral short hairpin RNA expression vectors for gene
Xing Zhong1, Chao Zhai, Dengxiang Yang
1Institute of Biochemistry and Molecular Biology, Hubei University, Wuhan 430062, Hubei Province, People's Republic of China.
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.
