Regular Plasmid Tet Inducible Gene Expr森道ession Vector

VectorBuilder’s regular plasm路術id Tet inducible gene expression v自草ector combines the regula相嗎r plasmid vector system wi司哥th the Tet inducible gene expression sy那費stem to help you achieve木她 simple transfection-based deliver慢資y of tetracycli站還ne-inducible gene expression 友河cassettes into mammalian南湖 cells.

The Tet-On induc老大ible system is a powerful tool to一又 control the timing of expression of紙暗 the gene(s) of interest (GOI) in mamm草暗alian cells. Our 員個Tet-On inducible gene expr雜一ession vectors are de能員signed to achieve低看 nearly complete silencing o呢了f a GOI in the abse大習nce of tetracycl花飛ine and its analogs (e.g. doxy文小cycline), and st購間rong, rapid expression近為 in response to the addition of te可章tracycline or o報件ne of its analogs (e.g. doxycyclin匠姐e). This is achieved我我 through a multicomponent system 車花which incorporates act學匠ive silencing by the tTS pr件業otein in the absence of tetracycli東舞ne and strong activ你對ation by the rtTA protein in 什他the presence of tetracycli場房ne. In the absence of tetracycline,數微 the tTS protein derived fro綠木m the fusion of TetR (Tet repressor pr如我otein) and KRAB-AB (the trans兒能criptional repressor domain o道司f Kid-1 protein) binds to t歌快he tetracycline公白-responsive eleme作海nt (TRE) promoter, leading to the a對民ctive suppression of gene transcrip相鐵tion. The rtTA protein,購體 on the other hand, deriv刀空ed from the fusion of a mu黑開tant Tet repressor 藍微and VP16 (the transcription activator小中 domain of virion protein 16 of he快離rpes simplex virus), binds t雨了o the TRE promoter to activate g拍信ene transcription only in the presen花睡ce of tetracycline.

While our regular plasmid Tet ind畫爸ucible gene expre校草ssion vector include著行s an inducible gen爸話e expression ca著白ssette consisting of the TRE promote信花r driving the user-s費湖elected GOI, the西吃 TRE binding regulator友黃y proteins tTS and rtTA have t說銀o be provided using a separat謝鐵e helper vector to achieve tetracyclin坐村e induced gene expr明長ession in the presence of tetracycline報報, while minimizing leaky expr年了ession in the absence of tetracycli照美ne. 

Delivering plasmid vectors in愛音to mammalian cells by conve秒哥ntional transfection is one of 呢這the most widely used procedures in遠文 biomedical research. While a河著 number of more sophisticated分熱 gene delivery ve紙長ctor systems have been developed間金 over the years such a多和s lentiviral vectors, aden關視ovirus vectors, AAV vectors紙學 and piggyBac, conventional plasmid t紅銀ransfection remains the wo機公rkhorse of gene de術頻livery in many la湖報bs. This is largely due to its t山我echnical simplicity as well 道公as good efficiency in a w筆南ide range of cell types. A key 錯水feature of transfection wi線技th regular plasm相畫id vectors is that it is transie體志nt, with only a v見作ery low fraction of cells stably integ影影rating the plasmid in the genom車話e (typically less than 1%).

For further information about場間 this vector system, please r頻拿efer to the papers below.

High-level expression:又錯 The TRE promoter can dr紙大ive very high levels年區 of expression of t電懂he GOI in its induced state.動但 Additionally, conven下兵tional transfection 他鐘of plasmids often results 就子in very high copy nu笑鄉mbers in cells (up謝都 to several thou空妹sand copies per cell).筆很 This can lead to ve的從ry high expression levels o相和f the gene(s) carried on the vec筆購tor.

Technical simplicity: Delivering plasmid vectors into c吧地ells by conventional t關服ransfection is technicall務讀y straightforward, and far刀友 easier than virus-based vectors whi黃長ch require the packag裡又ing of live virus.

Very large cargo space:外的 Our vector can acco照書mmodate ~30 kb o畫有f total DNA. The plasm海件id backbone only occu美答pies about 3 kb, leaving pl雪她enty of room to accommodate 討費the user's sequen紙議ce of interest.