Conditional knockdown of target gene expression by tetracycline regulated transcription of double strand RNA.
In vivo electroporation has served as an effective tool for the study of developmental biology. Here we report tetracycline inducible gene knockdown by electroporation. Our system consists of genome integration of a cassette encoding long double strand RNA (dsRNA) of a gene of interest by electroporation, transcription of which is assured by RNA polymerase II, and induction of transcription of dsRNA by tetracyclin. Long dsRNA decapped by ribozyme in the cassette and without poly A tail is processed into siRNA within nuclei. We could successfully induce knockdown of En2 and Coactosin by Dox administration.
Department of Molecular Neurobiology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
Xubin
X
Hou
Minoru
M
Omi
Hidekiyo
H
Harada
Shunsuke
S
Ishii
Yoshiko
Y
Takahashi
Harukazu
H
Nakamura
D000818:Animals; D002642:Chick Embryo; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics
D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D013752:Tetracycline