PiggyBac Promoter Tes妹地ting Vector (for In Vivo Promoter Tes可路ting)

This vector system is designed 文遠for efficient analysis of m計影ammalian promoters in mouse mo雨科dels utilizing the piggy是子Bac transposon system. Typically,術明 a putative promoter of i的師nterest is cloned into this ve對如ctor, upstream of a L北見acZ reporter gene and the res到坐ulting construct i話日s used to make transgenic志亮 mice. Expression of the LacZ 體秒reporter in transgenic e員新mbryos or adult mice can then b林如e used as a readout of promoter acti兒木vity. This vector syst去村em is useful fo內自r identifying promote吃線r elements, determining ti相聽ssue-specificity of promo喝歌ters, comparing p男商romoter variants, li信她neage-tracing and man鐵志y other applications.

Our piggyBac tra歌些nsposon-based vector systems are highly通愛 effective for inserting forei也學gn DNA into the host genom呢遠e of mammalian ce物山lls. This system is友視 technically simpl員物e, utilizing plasmids (rather than vir是商al transduction) to permanen長作tly integrate your gene(s) of 姐小interest into the host genome.

The piggyBac syste低鐘m contains two vectors, b習著oth engineered as E. coli plasmids. One船呢 vector, referred to as the h業問elper plasmid, encode算子s the transposase. The o訊飛ther vector, re校有ferred to as the t場國ransposon plasmid票服, contains two inverted t現吧erminal repeats (ITRs) bracketing the r男兵egion to be transposed. The pro視線moter sequence of服學 interest is clon一區ed into this region. When the he秒遠lper and transposon plasmids are雜雜 co-injected or co-transfec會動ted into target cells, the transpos近坐ase produced from the helper woul愛窗d recognize the two I書個TRs on the transposon嗎花, and insert the flanked re了金gion including the two科草 ITRs into the host genome. Inserti聽時on typically occurs at host chr到這omosomal sites that contain the 拍器TTAA sequence, which is duplica商時ted on the two flanks of the int村們egrated fragment a信都fter transposition.

PiggyBac is a Class II transposon, me視鐵aning that it moves in a cut-and-pa女少ste manner, hopping from place to pl來畫ace without leaving船票 copies behind. (In contr年銀ast, Class I transposons move in熱男 a copy-and-paste m好唱anner.) Because 綠男the helper plasmid is only tra廠去nsiently introduced into hos科校t cells, it will get lo日服st over time. With the 黃空loss of the helper plasmid,你大 the integration of t制女he transposon in the genome of host cel為明ls becomes permanent北但.

For further informat有近ion about this 雨自vector system, please refer to the p分高apers below.