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We narrowed to 237 results for: Mammalian expression plasmids

Showing: 41 - 60 of 237 results

  1. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors...Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid of 2017 is the lentivirus...envelope plasmid pMD2.G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid has been... 2017) and expresses the VSV-G envelope protein under the control of the powerful mammalian CMV promoter...Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances...the broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin...to check out the Trono Lab website. If you have plasmids you’d like to deposit with Addgene, be sure to...

  2. Protocol - Bacterial Transformation

    Type
    Protocol
    ...replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian cell expression... also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA DNA Ligation...that will help to maintain the plasmid without rearrangement of the plasmid DNA. Additionally, specific ...tube of plasmid DNA and just need to transform bacteria so that you can grow up more of the plasmid) you ...efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea to use ...Learn how to transform E. coli with your plasmid of interest. Protocols...introduced into a cell. Transformation of bacteria with plasmids is important not only for studies in bacteria ...

  3. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...that the most common expression system within the Addgene repository is mammalian, but the largest source...transfer of genes is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed...synthetic biology page Search for a plasmid Find Addgene's most popular plasmids ...been coming to the Addgene website in search of plasmids. Now, scientists are beginning to see Addgene ...Addgene as a large data set. Addgene has over 65,000 plasmids in the repository, each verified by sequencing...a distantly related organism, the gene may not express well because of these different codon usage biases...closely resembles that of the host organism, it may express protein more efficiently. To classify gene sequences...

  4. 27 Hot Plasmids from 2016

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    Blog Post
    ... Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry Mammalian Expression...lines expressing it. The Church lab fixed this issue by creating a new dCas9-VPR expression plasmid that...transfer vector for in-vivo expression and plasmids optimized for dCas9-VPR expression in drosophila and yeast...Cas9 nickase, and Cas9 activator expressing plasmids that can co-express 1-8 gRNAs. Ordon et al., 2016 tested...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...

  5. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...multiplexing; expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although...tracking, modulating splicing, and regulating expression of specifically targeted RNAs. Finally (and perhaps...COVID-19 (Joung, Ladha et al., 2020). Cas13a in mammalian cells The Zhang lab made the exciting discovery...discovery that some Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They...superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage with...at this finding through analyses of global RNA expression, cell stress, and RNA size distribution. When...27256883. PubMed Central PMCID: PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky,...

  6. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...Resources on Addgene.org Find Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR ...the techniques and the plasmids used to inactivate specific genes in mammalian cells. Despite the popularity...DNA designed in the lab. Specially constructed plasmids can be used to replace genes in yeast, mice, or...mind when choosing a knockout strategy! Knockout plasmids Homologous recombination is a mechanism to accurately...randomly integrated all or large portions of the plasmid. Deliver your construct to your target cells. After...Cre-mediated recombination.   Knockout/Knock-in plasmids Gene targeting methods also make it possible to... and insert GFP into the genome, we’d create a plasmid similar to the one shown below, where the sequence...

  7. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...using a plasmid with multiple, individual expression cassettes, having the genes expressed from the same... for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...cells that are expressing your gene of interest at high levels. Unlike vectors that express screenable or...multicistronic plasmids ensure that any cells that are positive for your marker should also be expressing your ...want to express multiple genes in the same cell. Although it is possible to drive co-expression by using... a portion of the plasmid is packaged for viral delivery, or the relative expression levels between two...with a plasmid that already has the multicistronic element and reporter cloned in. In these plasmids you ...

  8. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...organism, desired expression level, and experimental purpose. For mammalian lentiviral expression, we could use...reaction creates an expression clone with all of the components necessary for gene expression.     The ...(indicated here by plasmid color), allowing you to easily select for the expression clone. You will also...desired constructs: lentiviral expression of the human KRAS gene in mammalian cells. STEP 1: Generate an ...favorite gene! STEP 2: Generate an Expression Clone When making the expression clone, it is important to choose...clone to create the expression clone.    Figure 6: Generating an expression clone. The reaction...  STEP 3: Express your Gene of Interest! Be sure to verify the integrity of your expression clone via ...

  9. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ... is available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). ...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...Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 ... can be used to visualize RNA localization in mammalian cells. These tools consist of cobalamin-binding... the same number of fluorophores. You can find plasmids containing the cobalamin binding riboswitches ...

  10. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...tool of choice for a number of model organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish...Activity 35 - 45 °C 50 - 70 °C Genome Editing in Mammalian Cells? Yes Yes Longevity in Serum?* No Yes ...temperatures ranging from 25-80 °C. Activity is expressed as Kcleave (min-1). SpCas9 has maximum activity... experimental results. GeoCas9 RNP editing in mammalian cells Besides being able to take some heat, GeoCas9...GeoCas9 can introduce genomic edits in mammalian cells. Harrington et al tested GeoCas9’s editing skills...you want to give GeoCas9 a try, as always, the plasmids are available from Addgene. Happy CRISPRing! ...Background with Addgene’s CRISPR Guide Page Find CRISPR plasmids at Addgene ...

  11. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        New toolkit...tagging proteins with a 7kD degron called mAID and expressing the essential plant protein TIR1 in non-plant...was able to translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, ... the Kanemaki lab has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins ...previously difficult. Find SpCas9 and Target-AID-NG plasmids at Addgene! Nishimasu H et al., Science, 2018....

  12. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...types of yeast plasmids are defined below: Yeast Integrating plasmids (YIp): These plasmids lack an ORI...our first few Plasmids 101 posts, we focused mainly on the elements required for plasmid maintenance within... simple gene replacement/mutation. Yeast expression plasmids used in the lab typically contain all the...not only the number of plasmids found in each cell, but also whether the plasmid gets integrated into the...integration. Yeast Episomal plasmids (YEp): These are most similar to bacterial plasmids and are considered “...experiment. The marker provided by the plasmid may be expressed at higher than normal physiological levels...of marker gene expression is to use a partially defective promoter to drive expression of the selection...

  13. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...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 ...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...do many γ-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to...components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest..., which facilitate integration of the transfer plasmid sequences into the host genome. Typically, it is... genome upon viral transduction. Most transfer plasmids available from Addgene were derived from MoMLV...

  14. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...in-frame to the protein of interest for plasmid-based expression systems. Degron tags are one of many tools...be engineered or expressed beyond the tag and small molecule. It does work in mammalian cells, mice, and...on Addgene.org CRISPR Plasmids – Protein Tagging CRISPaint Gene Tagging Plasmids 101 eBook Resources ...Resources on the Addgene blog Plasmids 101: Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells ...the modulation of endogenously and exogenously expressed proteins. Here we will review what advantages ... tools to manipulate protein expression. These tags offer several advantages over the alternative systems...used to manipulate endogenously and exogenously expressed proteins. Thirdly, degrons and their drugs are...

  15. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ...website for plasmids that contain tinman or the mammalian homolog Nkx2.5. We also have one plasmid, Addgene... 1998). Interested in plasmids that contain the groucho gene? Check out plasmids deposited by the Shivdasani... several plasmids that contain SHH, you can find them all here! The most popular SHH plasmid however was...integrate particular genes into mammalian genomes and easily disrupt gene expression to study gene function. ...spaghetti. We have two plasmids in our inventory from the lab of Eric Wieschaus that express sqh.   cookie monster... to see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman...Drosophila Read our Plasmids 101 blog series Resources on Addgene.org Find plasmids for your research ...

  16. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...isolate individual plasmids, or express a gene contained in a plasmid. During the propagation and/or transformation...resistance DNA plasmids are traditionally grown in bacteria as a way to produce more of the plasmid, isolate...receive a copy of the plasmid or will lose it during division. These non-plasmid-containing bacteria can...the goal of your experiment — to grow, express, or isolate plasmids. The presence of an antibiotic resistance...selection, whereas other plasmids must carry both a bacterial and a mammalian antibiotic selection gene...bacteria that express the Tn5 element, but thankfully, most competent cells do not express this factor....Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog Plasmids 101: Antibiotic Resistance...

  17. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ...of interest has been cloned into an expression plasmid, the plasmid can be introduced into host cells, ...using synthetic genes that are typically expressed from a plasmid or from an integrated sequence in a stable...that the plasmids and sequences for antibodies are not shared. For instance, through open plasmid sharing...Bradbury et al., 2018). Hybridomas can lose gene expression of the mAb-encoding genes or fail to revive after...Recombinant monoclonal antibodies are encoded in plasmids and produce a single antibody recognizing one ...heavy chain genes of the antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an...identifying and cloning the genes for antibody expression. If starting from a protein sample (for instance...

  18. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...co-injection of the targeting vector and a Cas9 expression plasmid will insert the desired genetic tag and marker...as a universal donor plasmid. The CRISPaint toolkit includes several donor plasmids for protein tagging...collection of plasmids to fluorescently tag markers of cellular structures in mammalian cells. These plasmids...The Jorgensen Lab recently developed a modular plasmid assembly toolkit called SapTrap to improve CRISPR...accepts the sgRNA target sequence for U6 promoter expression and the second site accepts the homology arm ...repair template. SapTrap includes a prebuilt donor plasmid library containing several types of fluorescent...tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable marker, and...

  19. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in an inactive...bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p homolog, can also be used to monitor autophagy...yeast Rosella is targeted to the cytosol when expressed without a signal sequence and to the mitochondria...measurements of macroautophagy and mitophagy in mammalian cells using a single fluorescent biosensor. Scientific...Yoshimori, T., & Levine, B. (2010). Methods in Mammalian Autophagy Research. Cell,140(3), 313-326. doi:...Enzymology Autophagy: Lower Eukaryotes and Non-Mammalian Systems, Part A, 33-42. doi:10.1016/s0076-6879... Resources on Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors...

  20. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...on the Addgene Blog Plasmids for endogenous gene tagging in human cells Plasmids 101: Protein Tags Fluorescent... insertion of epitope tags at target genes in mammalian cells (Bressan et al. 2017; Liang et al. 2017)...from many pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis...Cas9 protein-based method for epitope tagging in mammalian stem cells. For easy scalability to the 96-well...Ribonucleoprotein (csRNP) delivery Epitope tagging of mammalian genes using CRISPR depends on successful delivery...directly purchased and there is no need to construct plasmid vectors or synthesize/purify IVT sgRNA in the lab... (although we could successfully knock-in non-expressed genes with similar efficiency, Dewari et al. 2018...
Showing: 41 - 60 of 237 results