...our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...development of future therapies. Our popular plasmid encoding a Cre dependent version of the DREADD hM3D is...are now available as AAV1. CRE-DOG constructs 69570 and 69571 together encode the N and C terminal segments...dependent Cre. The two segments come together in the presence of GFP to form a functional Cre enzyme that...both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available in AAV1. Plasmid Serotype...ChR2 for targeted expression DREADDs GFP dependent Cre and FLPo And more!  New lentiviral preps SARS-...protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the beginning...

...engineering efforts have increased the brightness of these genetically encoded light emitters (i.e. Takai...the most popular, most widely used genetically encoded fluorescent probe. Several factors contribute to... the first choice when selecting a genetically encoded fluorescent tag. There are, however, a number of...may alter diffusion rates. Smaller genetically encoded probes are based on cofactor binding proteins as...Both systems are a hybrid between a genetically encoded tag and exogenously added components. For a review...date, small probes that are entirely genetically encoded do not exist, but the development of non-natural...are made by connecting its DNA code to the cDNA encoding for another protein. Effectively, it equips the...

... involving two-plasmid or single-plasmid systems (Vo et al., 2020). In the two-plasmid system, an effector...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined... mini-transposon Transform plasmid(s) into cells and plate overnight Screen colonies phenotypically or...(V. cholerae INTEGRATE - hereafter VchINT) and encodes a Type I-F CRISPR-Cas system. The four major components...group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express the...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and ...

...) Interested in More Hot Plasmids from Addgene? Check out our Hot Plasmid page, where we highlight some...popular & newly available plasmids at Addgene. If you're interested in plasmids for in vivo imaging, check...for Neurobiology has been working on genetically encoded indicators of calcium and other small molecules...the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors Griesbeck’s recent contribution... including his zinc sensor and his genetically encoded magnesium sensor, the first of its kind. “I hope...Lindenburg et al. MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. 2013 Dec 2...large-scale functional screen in bacteria. Sensor variants were then refined by a secondary screen in rat hippocampal...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...ChiP-seq. Learn more about ENCODE Find the transcription factor tagging plasmids If you have a new CRISPR...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...post Read the paper in Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control...paper in Nature Methods Find the anti-CRISPR plasmids Tagging human transcription factors with GFP...

...control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid Serotype...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...biosensor AAV inventory New glutamate sensors Plasmid Serotype Name 106176  AAV1  pAAV.hSynapsin.SF-iGluSnFR.S72A...pAAV.hSynapsin.SF-iGluSnFR.S72A New calcium sensors Plasmid Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE...pAAV-hSynapsin1-FLEx-axon-GCaMP6s New voltage sensors Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...Sternson lab recently deposited PSAM AAV, which encode engineered ligand-gated ion channels that can be...of DREADDs / Chemogenetics AAVs New DREADDs Plasmid Serotype Name 75033  AAV1 pAAV CD68-hM4D(Gi)-...

...combines the Cre/lox system with optogenetics. Cre activity is controlled by red light by splitting Cre and fusing... Figure 4: The C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome... previous Cre/lox or drug-induced Cre systems (ex: tamoxifen). As the name suggests, CreLite combines ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...genetically encoded voltage indicator Article contributed by Shreya Vedantam Genetically encoded voltage ... dimerize, bringing each terminus of Cre together to restore Cre activity. As soon as red light or PCB...

... are not encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging...components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: ...γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (Vesicular...in trans by specific plasmids. This method, therefore, reduces the number of plasmids that are required ...γ-retroviruses self-inactivating (SIN)? They can be. Plasmids that encode SIN γ-retrovirus lack most of the U3 region...do many γ-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to..., which facilitate integration of the transfer plasmid sequences into the host genome. Typically, it is...

...deposited the resulting optogenetic plasmid tools which includes plasmids encoding a fusion between cPYP and tgRFP...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.          Listen to this episode of Hot Plasmids! Optogenetic...Grefen and Blatt, 2012). The 2in1 Plasmid Toolkit consists of 4 plasmid toolkits generated with 4 different... tgRFP (PLL7.0 tgRFP cPYP), and plasmids that encode the protein binders (which bind to either the light...targeting and editing experiments. Find the CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit ... from the El-Samad lab recently deposited plasmids encoding synthetic yeast transcription factors that...activation of gene expression. In addition to plasmids encoding these synthetic transcription factors, the...the El-Samad lab has deposited plasmids encoding fluorescent reporters and other proteins controlled by...sign up here. No time to read? Listen to our Hot Plasmids Podcast! TRIM-Away: Targeted endogenous protein...

...is split into two plasmids: one encoding Rev and one encoding Gag and Pol, increasing the number of recombination...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 long... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ... Lentiviral Plasmid FAQs. Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...

...on the plate. For example, if your plasmid contains a gene encoding a blue chromoprotein, after transforming...colonies to determine which cells have taken up the plasmid. Of course, other follow up methods like sequencing... chromoprotein sequences into E. coli BioBrick plasmids (meffRed, eforRed, asPink, spisPink, scOrange,...chromoproteins. In some cases, cells containing high-copy plasmid-borne chromoproteins lost color overnight in liquid...problem. There is no single “ideal” chromoprotein plasmid that combines the most desirable features: intense...Chromoproteins for allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus...sequential homologous recombination events: The plasmid carrying the desired genetic modification integrates...

...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well as sequences...Psi). Because the packaging plasmids lacked the psi signal, the genes encoding the structural proteins could...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector genome which was composed...expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...

...instance, Lifeact-mTurquoise2 (Plasmid #36201) or mScarlet-tubulin (Plasmid #85045). We often use restriction... organisms. Both components of the chimera are encoded by DNA. Since researchers can generate almost any...Aequorea victoria and its variants are genetically encoded fluorescent probes. One of the limitations is the...over-expression artifacts The introduction of DNA that encodes a fusion protein adds protein to the existing pool...advisable to first express the fusion protein from a plasmid-based system to assess whether the chimera is still... develops, characterizes, and uses genetically encoded fluorescent probes. You can follow him on twitter...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...

...labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your favorite ...ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine (Cys4) peptide tag could be used ...These widely used systems consist of a genetically encoded enzyme variant tag that reacts specifically and...paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule binding motif...trade-offs between the size of the genetically encoded tag, the speed and selectivity of the fluorophore... Generate Fluorescent Protein Fusions Request Plasmids from the Michael Davidson Collection Find Fluorescent...overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores, compromising the photon...

...pool and extract plasmid DNA directly from the colonies. Transform the resultant plasmid DNA into E. coli...Assuming they expressed well, the proteins encoded by the plasmids in the library will remain attached to ...pinpoint the genes within a sequenced genome that encode for functional cell-binding proteins in the (all-too-common...DNA bases.  Ligate and transform your resultant plasmid libraries into chemically competent E. coli and... you’re using a pET vector) and antibiotic for plasmid selection. Remove air bubbles between the membrane...the original plate to identify which phage gene encodes that protein! First, block your membranes with ...Exploring the interactions between bacteriophage-encoded glycan binding proteins and carbohydrates." Current...

...interest endogenously and transfect them with a plasmid encoding your protein of interest. Then you should ... protein in a Western blot. You acquire a plasmid encoding your protein of interest, known as Gene of...parallel, you transfect cells with another plasmid not encoding this protein, which we will call Negative...blog post!  Resources on Addgene.org Find plasmids encoding antibodies at Addgene Stay tuned for ready-to-use...band from the cells transfected with the other plasmid. Your antibody works! Multiple (Independent) Antibodies...Designing a recombinant validation experiment? Find plasmids here!  References Baker M (2015) Reproducibility...IWGAV five proposed antibody validation pillars. Created with BioRender.com.     Orthogonal Strategies...

...transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell...) alone at 1.1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...

...the Cre-dependent SPH construct for generating the SPH mouse. (B) AAV plasmids used to express CRE under... of Cre-dependent mouse lines for overexpressing genes within discrete cell populations makes Cre-dependent...Constitutive or inducible Cre recombinases can be introduced by AAV or by mating with Cre-expressing mice. ...neuronal populations.  The Yang Lab’s AAV plasmids used to express Cre under neuron-specific promoters, including...comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ... sensitive applications such as CRISPR screening. The plasmids used by the Qi Lab for this particular ...genes can be relatively laborious because multiple plasmids/viruses are needed to deliver all the protein ...

...molecular biology reference Find Cre-lox plasmids at Addgene Browse plasmids at Addgene ...cells with an inactive opsin gene; in Cre expressing cells, however, Cre induces the recombination of the ... about FLEx vectors Download our Plasmids 101 eBook Read our plasmids blog posts Resources on the Addgene...(Ernst et al., 2014). Microbial rhodopsins are encoded by opsin genes, and include bacteriorhodopsins,...researchers inject genetically modified viruses encoding microbial rhodopsins into specific cerebral regions...achieved through site-specific recombinases such as Cre or Flp that induce DNA recombination at defined recognition...loxP and lox511) in opposite orientations. Since Cre does not cause recombination between mismatched recognition...