shRNA Expression Plasmids Generated by a Novel Method Efficiently Induce Gene-Specific Knockdown in a Silkworm Cell Line |
| |
Authors: | Hiromitsu Tanaka Kosuke Fujita Aki Sagisaka Kazuya Tomimoto Shigeo Imanishi Minoru Yamakawa |
| |
Institution: | Innate Immunity Research Unit, National Institute of Agrobiological Sciences, Ibaraki, 305-8634, Japan. htanaka1@affrc.go.jp |
| |
Abstract: | RNAi knockdown by using shRNA expression plasmids is widely used to determine the function of individual genes in mammals. Here we developed a simple method to create an IR DNA in a U6 small nuclear RNA promoter-based parent vector using a single-stranded IR DNA with short hairpin structure and Bst DNA polymerase. Furthermore, we demonstrated that the shRNA expression plasmids constructed by our method effectively induced target-specific RNAi in the silkworm cell line. We also found that sequence preference in the silkworm cell line was much lower than in mammalian cells and shRNA-induced RNAi was influenced by the length of the stem region. |
| |
Keywords: | RNA interference Bombyx mori Short hairpin RNA U6 promoter Bst DNA polymerase |
本文献已被 PubMed SpringerLink 等数据库收录! |