title: Generation of inducible shRNAi human embryonic stem cell lines.
abstract: This unit describes the generation of tetracycline-inducible short hairpin RNA interference (shRNAi) human embryonic stem cell (hESC) lines. Using this vector-based approach enables stable and long-term expression of target hairpins under the control of doxycycline/tetracycline. Target degradation can be controlled in both a dose- and time-dependent manner that can even be switched off, depending upon the particular requirements of the study.
authors.affiliation: Institute of Medical Biology, ASTAR, Singapore.
authors.forename: Stuart
authors.initials: S
authors.lastname: Avery
mesh_terms: D000818:Animals; D002021:Buffers; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D015536:Down-Regulation; D004318:Doxycycline / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009841:Oligonucleotides / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D002021:Buffers; D009841:Oligonucleotides; D034741:RNA, Small Interfering; D004318:Doxycycline