...calcium.”  Primary Vector Type The vector type should describe how your plasmid is intended to be used... such as lentiviral, retroviral, and AAV plasmids, choose the NEB Stable strain. For plasmids containing...deposits with more than 10 plasmids is to use our deposit spreadsheet?  Our plasmid deposit process can be... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...spreadsheet and copy and paste your plasmid data directly into our file, one plasmid per row, as outlined by the... Data Field Description Plasmid Name Use a descriptive plasmid name and match the name as it ...application of the plasmid and will help a scientist outside your field understand if this plasmid will work for...

...subset of the new plasmids and viral preps 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...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...PAM. Get the plasmids at Addgene The Human GlycoGene CRISPR Library is a new lentiviral CRISPR library...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory...fluorescence from other regions of the cell. By combining vectors expressing different fluorescent proteins targeting...

...use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the...TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector containing...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector. The vector is 8901 base...Protocols pLKO.1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006...pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA...

...strains and lentiviral expression vectors (Table 1). The four systems Name Function Plasmid Transduction...Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells...deposit also includes pXPR_071, a plasmid that leads to higher titer lentivirus preparations than the first ...first generation CHIME (pXPR_053). Find X-CHIME plasmids here!   Marty LaFleur is a Postdoctoral Fellow in...

...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...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...on earlier evidence showing that a CRISPR/Cas9 plasmid could be injected directly into mice for genome...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames...which all but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker...

...the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell ...physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput screening...comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...Neuron-optimized viral vectors Alternatively, Cas9 activators can be delivered to the CNS by lentivirus, which has.... This dual lentivirus system pairs ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific...including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches. Here...genes can be relatively laborious because multiple plasmids/viruses are needed to deliver all the protein ...

...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...of producing infectious virus (similar to lentiviral vectors) because they do not encode all of the necessary...encoded on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome...four viral genes in the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... 2 μg minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...Minigenome System). From a more technical standpoint, a plasmid with all four genes and necessary control elements...

... the pOSIP (one-step integration plasmid) series of plasmids, vectors that convey both the sequence to... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...These vectors and sgRNA libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based... existing transposon and vector systems and creating an all-synthetic vector that included only the elements...see our CRISPR/Cas Plasmids: Pooled Libraries webpage or read our blog post,Lentiviral CRISPR Libraries ...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite...

... the type of vector that you will need. You will also have to make sure that your plasmid has been incorporated...fusion protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3, comes...will need accessory packaging vectors to generate the virus. Bacterial vectors This system is great for overexpressing...cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope...folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your applications...reading frame (ORF) of interest –  try pRS plasmid shuttle vectors. You may prefer the more traditional approach...conduct your custom experiment. Fly vectors The vast majority of fly vectors are based on p-element, random...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...with GeneWeld vectors GeneWeld from Jeffrey Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins...that targets the donor vector sequence. Homology arms that flank the donor plasmid cargo are complementary...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... tools for your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/...cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting requires two sgRNAS: one ... Read the paper in bioRxiv Find the GeneWeld vector kit CasX, a small Cas protein, is active in E....

...A key feature of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...robustness of the system and utility of the pRDA_526 vector. Moreover, the screen identified putative novel...FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction of the model antigen,...to study antigen-specific responses. Find FITS plasmids here! Marty LaFleur is a Postdoctoral Fellow in...

...that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic resistance...an integrated lentiviral vector. Protocols...expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term protein expression ...transfection approaches, generating cell lines using lentiviral vectors enables long-term protein expression studies...with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries...Stable Cell Lines with Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols...

...create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ... routes in various conditions. The plasmids and lentiviral vectors encoding these proteins can also be...amazing plasmid technologies developed by our community of depositors. With over 40,000 plasmids avaliable...amazing new plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool ...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...

...treatment. The library consists of a pool of lentiviral vectors with roughly 73 million semi-random, 30 bp...Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources...individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and ...optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later, Bhang says, “When...infection. When using the ClonTracer system, lentiviruses containing these barcodes infect the target ...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...components to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro...cytoplasm and can be fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4.... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...

...from a lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol... 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume 500 μL *Plasmid concentrations and ... each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid DNA purification...endotoxin removal step. For high quality plasmid DNA, the plasmids should also be propagated in an endonuclease... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...Viral Vector Guides Mol Bio Protocols Introduction This protocol can be used to produce lentivirus from...~20 h. Prepare a mixture of the 3 transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish...

...2016, Addgene has shipped thousands of AAV and lentiviral preps to scientists around the world. In the ...gradient ultracentrifugation This video goes over AAV vector purification using an iodixanol column gradient...instructions and calculations on how to prepare a plasmid stock used to generate a standard curve, visit ...

...Addgene.org Find Fluorescent Protein Plasmids Find Plasmids for FRET Find Plasmids for Subcellular Localization...would be successful and the importance of lentiviral vectors in ensuring that success Chapter 6 - Developing...organization (now with more than 50% international plasmid distribution!) Chapter 8 - Opportunities to create...

...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...about a recent plasmid deposit please sign up here.   Listen this episode of Hot Plasmids! Light controls...in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius A, et...like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and Cas9...Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 ... the same number of fluorophores. You can find plasmids containing the cobalamin binding riboswitches ...