Regular Plasmid Tet 數錯Regulatory Protein Expression V的鄉ector

VectorBuilder’s regular plasmid Te麗見t regulatory protein expression ve山醫ctor can be used fo制南r simple transfe工山ction-based delivery of Tet regula歌電tory protein(s) (tTS, rtTA,在愛 etc.) into mammalian cells to hel東體p you achieve tetracycline-inducible 視山expression of target genes placed downs黑我tream of a tetra妹做cycline responsive-element (TRE) 家算promoter.

The Tet-On inducible syste麗到m is a powerful tool to control 聽器the timing of expression of the gen兒愛e(s) of interes城街t (GOI) in mammalian cells. Our Te器年t-On inducible gene expre麗的ssion vectors are designe志月d to achieve nearly complete silencin現外g of a GOI in the absence of tetrac件道ycline and its an民腦alogs (e.g. doxycycline又雜), and strong, rapid expression in res是討ponse to the addition of tetra少花cycline or one of its a中你nalogs (e.g. doxycycline). This is ac說農hieved through a multic城費omponent system which incorpor友器ates active silencing by the tTS pro文外tein in the absence of tetracycli林視ne and strong activation by th得科e rtTA protein in the presen花說ce of tetracycline. In the 黃短absence of tetracycline, the 東月tTS protein derived放新 from the fusion of章坐 TetR (Tet repressor protein) 白到and KRAB-AB (the transcriptional re呢很pressor domain of明森 Kid-1 protein) 內讀binds to the TRE promoter, leading 新哥to the active suppression of gene tran報拿scription. The rt分說TA protein, on the商科 other hand, derived from the 海在fusion of a mutant Tet repressor 票跳and VP16 (the transcr火些iption activator doma一裡in of virion protein 16 of分日 herpes simplex virus), binds to the 你鄉TRE promoter to activate gene transcri你西ption only in the presence of tetracycl關家ine.

While our regular pl短計asmid Tet regula微機tory protein expression ve老土ctor includes an ex很金pression cassette consi長友sting of the Tet regulatory protei工街n(s) driven by a user-select紙下ed promoter, the GOI driven行員 by the TRE promoter術用 must be provided us窗上ing a separate helper vector t吧如o achieve tetracycline in西拍duced gene expression in the 劇從presence of tetracycline, whi見報le minimizing leaky空家 expression in the ab友業sence of tetracycline.

Delivering plasmid vectors into拍術 mammalian cells by 就站conventional transfection is one 件老of the most widely used procedures i從離n biomedical research見得. While a number of more soph喝我isticated gene delivery vector systems對街 have been developed厭黑 over the years such as lentiviral vect什舞ors, adenovirus vector麗熱s, AAV vectors and piggyBac, convention物筆al plasmid transfection r問城emains the workhorse o國關f gene delivery in many明歌 labs. This is largely due to its techn公做ical simplicity區算 as well as good effic中吃iency in a wide range of cell types. A 新他key feature of tra拍討nsfection with regular plasmid為文 vectors is that it is transient, w人又ith only a very low道火 fraction of cells stably inte兒熱grating the plasmid in the genome (typ鐵白ically less than 1%).

For further information about數弟 this vector system, 什黑please refer to the papers below.