...and constitutive promoter. Use this for doxycycline inducible (tet-off) gene expression! We've also added...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share...share the 20 additions we've made to our viral vectors repository since March.  Addgene is now offering ... precision.   Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category... multiple serotypes Looking for more viral vectors? Browse the full viral services collection or check... check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the...below to keep up with the latest in Addgene's viral vector services and resources!   This blog post was written...

... set of vectors for inducible expression (pSBtet derivatives). The constitutive and inducible expression...efficiencies of integrating viral vectors. New Sleeping Beauty transposase vectors Even more recently, Eric ...developed two new types of vectors for the SB transposase system - one set of vectors for robust constitutive... inducible vectors...cells’ defense mechanisms. The use of non-viral vectors, like transposons, avoids many, though not all,...expression vectors were developed into 16 variants with different selection markers (blasticidin, neomycin, ... protein reporters (GFP, BFP, RFP, no FP). The vectors each contain two SfiI restriction sites with different...

...gene expression by administering an inducing agent like doxycycline (Favre et al., 2002). They are useful...experiment. Some inducible constructs can exhibit leaky expression in the absence of the inducing drug, so be...on the Addgene blog Viral Vectors 101: An Introduction to AAV Viral Vectors 101: The AAV Retrograde Serotype...Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst...testing, and optimizing three key variables of an AAV vector: serotype, promoter, and titer.     Figure...efficacy. You’ll therefore need to test potential AAV vectors in your specific experimental system, rather than...it can be ubiquitous, cell-type selective, or inducible.  Ubiquitous promoters  These often-strong promoters...

...common drug used to induce the expression of Cre is tamoxifen. In tamoxifen-inducible Cre (Cre-ER) systems...include tetracycline or doxycycline mediated activation of Cre to either induce or repress Cre expression...Mice can be engineered by inserting a targeting vector containing the gene of interest into a zygote, ..., knockout mice, and mice with conditional or inducible gene expression. Transgenic and knockin mice As...microinjection of DNA or infection with a viral vector containing the gene construct can be used to insert...generated via homologous recombination. A targeting vector is electroporated into embryonic stem cells and...into a female animal for gestation. Nucleases that induce double-stranded DNA breaks, such as ZFN and TALEN...

...  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David Root lab is...capable of inducible expression of shRNA in target cells by the addition of tetracycline/doxycycline to the...opening our UK office in 2014, to starting our viral vector service in 2016...we’re excited to help scientists... be used with 2nd or 3rd generation lentiviral vectors and an envelope expressing plasmid, such as our...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver Cas9...your gRNA of interest into mammalian cells. This vector was the top requested plasmid of 2015. What makes... This plasmid from the Zhang lab is a mammalian vector that expresses both SpCas9 and your gRNAs and was...

...cycle, a transfer vector with an inducible promoter (see below) should be used. Inducible lentiviral transfer...transfer vectors The most widely used inducible lentiviral vector system is the tetracycline (Tet)-regulated...your experiment using the lentiviral vector system. Viral vectors have been increasingly popular in fundamental...primary cells. Amongst the different vectors used, lentiviral vector constructs have proven very useful ...fluorescent sensors. Retroviral vector background Until the development of viral vectors, gene transfer into mammalian...un-transfectable cells, labs use viral vectors and particularly retroviral vectors because they target a broader...attractive choice for a gene transfer vector. Lentiviral vectors Although retroviruses are still in use...

... repressor in hPSCs is also driven by a doxycycline inducible promoter. The gRNA piggyBac plasmids include...pMAK backbone by modifying pMAL-T-Avi-His/BirA, a vector commonly used for in vivo biotinylation via BirA... and results in uniform NB biotinylation. Once induced, the NBs could be easily purified by one-step immobilized...Institute developed fluorescently labeled piggyBac vectors for uniform and sustained expression of single ... dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into genomic DNA with the help of the... T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate the nanobodies, the authors used...

... of Gateway cloning vectors Vector type  Vector features Purpose Donor vector attP sites for recombination... we could use a vector like pLenti CMV Puro DEST (w118-1) or the doxycycline-inducible pLIX_403. The chosen...sequences into multiple vector systems. With the appropriate entry and destination vectors, one can use Gateway...ccdB gene is present in the donor vectors and the destination vectors prior to recombination, and it is...donor vector.   Method B: TOPO-cloning of the desired insert into an attL-entry-TOPO vector. TOPO...Gateway vector to produce the desired expression clone. This is possible thanks to the Gateway vectors’ design...sites creates attL and attR sites.   Gateway vectors contain modified versions of the att sites so that...

...expression vectors with various promoters suitable for in vivo expression, and/or produce vectors to make...prototrophic strains. The vectors were based on the original EasyClone vectors (Jensen et al 2013) allow...donor vectors containing mAID and either Neo or Hygro resistance markers with CRISPR/Cas vectors targeting...control of the TRE promoter, which allows for doxycycline-inducible expression. The end result is genomic integration...screen for genes involved in West Nile Virus (WNV) induced cell death. The Wu lab library consists of 77,406... that positively regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3... interest and cloned into provided destination vectors using low-cost, one-pot golden gate cloning that...

... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers from Table...homology between the vector and gene primer for SLIC cloning. Amplify the pCoofy vector and the sequence ...recombination in SLIC cloning to form the desired vector. The pCoofy vectors contain the ccdB cassette, which is not...