...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... interest and cloned into provided destination vectors using low-cost, one-pot golden gate cloning that... Jacks Lab constructed several GMAP-compatible vectors for a wide variety of biological applications. ... problem, Csaba Pál’s lab has created a set of vectors that allows one to use the MAGE method in unmodified...clinically relevant bacterial species. pORTMAGE vectors open new horizons to modify genomes in a broad ...

...into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type IIS restriction...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...create lentiviral vectors expressing wt humanized Cas9 and up to three gRNAs. Entry vectors containing the...wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing other...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...assembly of multiple components into a destination vector. The first step in CRISPR/Cas9 Golden Gate multiplexing...to allow for ordered assembly into a destination vector that expresses Cas9. This is illustrated in the...

...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with the...MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and a collection of building blocks...gene expression studies, which are evinced by the Tet system and visualization of single molecules via .... Additionally, by using an adapted destination vector with a heterologous promoter outside of the Multisite...

...Mice can be engineered by inserting a targeting vector containing the gene of interest into a zygote, ...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...engineer knockin mice. Co-injection of a targeting vector containing homologous ends to the breakpoint promotes...promotes homologous recombination of the targeting vector. CRISPR/Cas9-based systems have been developed ...the expression of a particular gene. Targeting vectors used to create knockout mice include a reporter...standing technique is the injection of a targeting vector into embryonic stem cells, which are then injected...

...building block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial...research purposes, retroviral vectors have been developed into ‘split-vector systems’ by separating viral...and expression of these vectors when used in experiments. Retroviral vectors make up a large toolbox ...Gamma-retroviral vs. lentiviral vector systems Gamma-retroviral vectors have a relatively high transduction... many more useful viral vectors available at Addgene: Follow our viral vector blog posts for updates on...tips for using retroviral vectors in your experiments. Gamma-retroviral vectors Popular gamma-Retroviral...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...

... a plant cloning kit that contains a series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs...is available in serotype AAV1. The GCaMP8 viral vectors from the Janelia Farm GENIE project have arrived...arrived! We now offer several versions of GCaMP8 vectors in serotypes AAV1 and AAV5. AAV9 preps and more AAV1...projections and putative pre-synaptic terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from...project. We now have over 40 INTRSECT 2.0 project vectors available as ready-to-use viral preps with more...on the way. S5E2 regulatory element containing vectors developed in the Jordane Dimidschsteind lab provide... interneurons. We now provide ready-to-use AAV vectors expressing GCaMP6f with this promoter. Image calcium...

...hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation and inhibition by using both...candidates for your experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated...photoswitchable tethered ligands (PTL), and photoswitchable, orthogonal, remotely-tethered ligands (PORTLs... secondary signaling pathways. Photoswitchable tethered ligands hold the ligand in close proximity to ...

...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production...acids. This conjunction of vectors and exosome was given the name vexosomes (vector-exosomes), exosome AAVs... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...production, assembled AAV vector particles accumulate inside the producer cells and purification protocols...purification steps. It’s also possible for AAV vectors to be spontaneously released from the producer ...Harvard Medical School took a closer look at the AAV vectors that are released from cells during production ...advantages over the conventionally purified AAV vectors. They found that they have a higher transduction...

...A newly established all-in-one vector construction system for CRISPR/Cas9-mediated multiplex genome engineering...systematically established way of making an all-in-one vector for multiplex genome engineering.” Now, they’...ease and efficiency for all-in-one CRISPR/Cas9 vectors expressing up to seven guide RNAs (gRNAs). Sakuma...Schematic overview of the all-in-one CRISPR/Cas9 vector construction system for multiplex genome engineering...expression cassettes of the gRNAs into a single vector in tandem. They went on to show that their CRISPR...CRISPR/Cas9-nuclease and CRISPR/Cas9-nickase vectors work successfully to target multiple sites in cultured...it has not yet been demonstrated, they say the vectors are also likely to work in animal embryos based...

...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...pSBbi derivatives) and one set of vectors for inducible expression (pSBtet derivatives). The constitutive...cells’ defense mechanisms. The use of non-viral vectors, like transposons, avoids many, though not all,...constitutive and inducible expression vectors were developed into 16 variants with different selection markers (... protein reporters (GFP, BFP, RFP, no FP). The vectors each contain two SfiI restriction sites with different... and gene disruptions without the use of viral vectors. In 2009, it was named Molecule of the Year. Additionally...

...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...check for this in your system and then test each vector before starting your experiments (Rolland et al...sure to test for this when validating your viral vector. Titer Once you have selected potential serotypes...off-target effects (e.g. recombinase-dependent vectors) increases with the amount of virus you add, so...

... into and storage of lentiviral and retroviral vectors or cloning or repeated sequences with the potential...storage of unstable DNA and lentiviral/retroviral vectors. Used by Addgene for pooled library amplification...Replication The Promoter Region – Let's Go! Mammalian Vectors Looking for More Plasmids 101 Articles?  Find ...Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 tetR F'[proAB lacIqZΔM15 Tn10(TetR Amy CmR)]  Note: Inactivating ... streptomycin Tn10   Confers resistance to tetracycline Additionally, Table 2 provides a quick reference...

...AAV1 pAAV-ihSyn1-DIO-tTA Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have released in the...AAV pEF1a-DIO-FLPo-WPRE-hGHp Other new viral vectors New optogenetics inhibitors Plasmid Serotype...for order please email [email protected] with the vector name and serotype you’d like to be notified about...  AAV5 ppAAV-CKIIa-stGtACR1-FusionRed New tetracycline transactivators Plasmid Serotype Name 99120...

...communication. Viral vectors and the nervous system With the advent of viral vector technology, new methods... at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have been widely used...genome is a single-stranded DNA molecule and AAV vectors are replication deficient (i.e. they cannot easily... low immunogenicity and cellular toxicity. AAV vectors can be equipped with strong promoters for high ... is often made Cre-dependent. If Cre-dependent vectors are used in transgenic animals that express Cre...that also express Cre. Another advantage of AAV vectors is that there are well-established protocols for...life cycle. Using AAV for neuronal tracing AAV vectors are the first choice of many neuroscientists, with...

...cloning vectors such as pBR322, pACYC, and pUC were developed to provide higher copy number vectors that ...successful, pSC101 became the first plasmid cloning vector and molecular biology was never the same. Over ...the "Plasmids that Glow" and "Viral Expression Vectors" chapters, and more pictures and pro tips.  Happy...experiment. Dr. Cohen and colleagues treated a tetracycline resistant plasmid, pSC101, and a newly developed...

...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...indicate type in “Vector Type Other” column. For example, you may enter “donor template vector”. Cloning...plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids ...nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline, combinations of the above, or other. High...

...plant We developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...genome. We constructed three versions of the vector (pKABE6.3, pKABE7.9, and pKABE7.10) so that the user...are better suited (Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA... genomic site simultaneously.     These binary vectors could be used to generate stable genome edited ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast ...et al., 2016) invented CRISPR-base editors by tethering cytosine deaminase with Cas9 nickase (nCas9). ...

...Recently the Vellai lab has created an improved RNAi vector that leads to significantly stronger phenotypes...for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted...complexes with LacZ𝜔 and forms an active LacZ tetramer.  This tetramer cleaves chlorophenol red-b-D-galactopyranoside...

... construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many...Shetty RP, Endy D, Knight TF. Engineering BioBrick vectors from BioBrick parts. Journal of biological engineering... fluorescent protein were fused in-frame to a tethering partner, which directed localization of the fluorophore...Construct Fluorophore Excitation/emission maxima Tethering partner Subcellular localization 1 TagBFP 399nm...

...lab developed new fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of ... RNA-guided methylcytosine editing by TET1 by co-delivering TET1 and protein factors for linking methylcytosine...., 2019. Fusing epigenetic modifiers such as TET1 with inactive Cas enzymes can alter gene expression...