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We narrowed to 162 results for: Mammalian expression plasmids

Showing: 21 - 40 of 162 results

  1. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...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...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'...

  2. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...designed to create plasmids for fluorescence imaging or flow cytometry applications in mammalian cells. The modules...Gibson Assembly Learn About Other Plasmid Cloning Techniques Plasmids 101: Green Fluorescent Protein Resources...Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid Cloning Protocols References 1....combination of different genetic sequences into plasmid DNA. In high throughput cloning techniques, although...genetic elements may differ (e.g., a set of various mammalian promoters), the same cloning procedure can be ...localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu, 2015...enhancers, polyadenylation signals, inducible gene-expression sequences, and more. By combining these components...

  3. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...couple termination with polyadenlyation. Mammalian expression plasmids are primarily used to create mRNA and...Plasmids designed to express genes in a given host cell type are generally broken down into two broad...Download our Plasmids 101 eBook to get all the background you need to start working with plasmids! What are...terminators are not usually employed in plasmid-based expression systems, so these will not be detailed...at the end.  Nearly all common bacterial expression plasmids use Rho-independent terminators, which include...Promoter Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids 101 Posts Resources...Browse our Plasmid Reference Pages Just Starting Work in a New Field? Check out Our Plasmid Collections...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...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 ... mediated genome modification and gene expression. The plasmids associated with this article can be obtained...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase... of the integrases is supplied on its own expression plasmid or in various combinations, while the cognate...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...Fortunately, a collection of plasmids comprising the multiple lentiviral expression (MuLE) system from the ...

  5. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    Published
    May 12, 2015, 1:32 p.m.
    .... When expressed in neuronal cells, each line showed broad recombinase-dependent RFP expression. Even ....1111704108. Epub 2011 Aug 9. PubMed. Find the Plasmids @Addgene: Plasmids for KD, R, B2, and B3 recombinases from...problem is especially stark in Drosophila, where expression of Cre from the standard UAS/GAL4 system is toxic...Even at low expression levels, recombination by KD, B2 and B3 resulted in complete or near-complete excision...two of these recombinases were also active in mammalian cells. In experiments, B3 and KD were shown to... and DNA damage induced by Cre recombinase in mammalian cells. Loonstra A, Voojis M, Beverloo HB, Al Allak.../s004270100167. PubMed. Cryptic loxP sites in mammalian genomes: genome-wide distribution and relevance...

  6. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...Addgene includes bacterial and mammalian mCyRFP1 or CyRFP1 expression plasmids, FRET constructs, and CaMKII...Species Bacterial Expression Construct SpCas9 Inhibition (E. coli) Mammalian Expression Construct SpCas9...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...articles provide a synopsis of a plasmid or group of plasmids' functions and applications. We hope that... you to find and use the plasmids you need. You can find all the hot plasmids from 2017 below. With over...over 50,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...

  7. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...brightness When a dish of mammalian cells is transiently transfected with a plasmid containing a fluorescent...individual mammalian cells within the dish. This is caused by the stochastic nature of plasmid uptake by...et al, 2012, Bindels et al, 2017). The plasmid for co-expressing mTurquoise2 with mVenus as a reference...variation, a reference fluorescent protein that is expressed at an equal level (Goedhart et al, 2011) can be...Another way to dodge cellular variation observed in mammalian cells is to tag an endogenous gene. By imaging...Mouridi et al (2017) and Heppert et al (2016). In mammalian cells this would be possible with CRISPR/Cas-based... The practical brightness assay (employing co-expressing fluorescent proteins or using endogenous tagged...

  8. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast, and mammalian. These...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...interacts with human cells. Addgene’s mammalian target plasmids express these and other targets such as TMPRSS2...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...spike protein to improve expression and pseudotyping efficiency. This plasmid was later used by another...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from...

  9. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...cost-effective and faster-growing than mammalian cells, but mammalian cells can perform human-like post-translational...transfect your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell... great collection of ready-to-use rAbs and rAb plasmids!). Since recombinant antibodies conveniently allow... allow for unlimited production, reliable expression, and easy distribution as DNA (Trimmer, 2020), you...can take place in bacterial, yeast, plant, or mammalian cells. Each of these systems has their advantages...bonds (Frenzel et. al., 2013). If you’re using a mammalian production system, cell lines typically used include...antibodies can be produced through a single or dual-plasmid transfection, depending on whether the heavy and...

  10. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...  Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with four being...Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after Golden ...Golden Gate Assembly Multiplex Plasmids: These plasmids allow expression of 2-5 gRNAs in zebrafish. Custom...on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll use mammalian plasmids...Why use multiplexed gRNAs? By expressing multiple gRNAs on the same plasmid, you’ll make sure that each ...receive is: can I express more than one gRNA from a single promoter using a plasmid like pX330? Unfortunately... you’re working in Drosophila, a two-gRNA expressing plasmid is available from the Bullock lab, and gRNAs...

  11. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ...Cell Culture Cloning Mammalian Cells with the Agarose Method Plasmids 101: Mammalian Vectors Resources ....org Find mammalian plasmids for your research Find viral vectors for infecting mammalian cells ...cross-contamination can be identified. Even if some mammalian cells lines have similar morphologies, being familiar...)). Blood pH normally lies around 7.35-7.4 so mammalian cells are sensitive to pH. Anything between orange...LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced perturbation...cytoskeletal disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied...

  12. Antibodies 101: Affinity Tags

    Type
    Blog Post
    Published
    Sept. 19, 2023, 1 p.m.
    ... single protein is expressed, with the intent of purification, it will be expressed along with all other...cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually isolate...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and...more straightforward and of a higher purity than mammalian purifications due to the presence of more naturally...

  13. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ... Mammalian Transposon pRDA_757 (Piggybac; constitutive expression) Mammalian Plasmid pRDA...restriction digest, and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just ...correctly banded clones for plasmid sequencing and found 98% of the plasmids were perfectly correct. Because...Vector Mammalian Lentivirus pRDA_512 (RSV-driven; lentiGuide-based) Mammalian Lentivirus... over 512) Mammalian Lentivirus pRDA_789 (CMV-driven; CROPseq) Mammalian AAV pRDA_889... Organism (Mammalian or drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination...the assembled plasmid. It shows you each component, with the ID of the component plasmid, component name...

  14. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...restoring expression in the liver.  Read the paper in Nature Biotechnology Find the PEDAR plasmids   Additional...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...packaged into AAV particles for delivery into mammalian cells, especially when fused to effector proteins...for such AAV packaging, but their activity in mammalian systems has been in question. Three different ...this problem. A miniature CRISPR-Cas system for mammalian use Stanley Qi’s lab has developed a miniature...

  15. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...transforming both fusion plasmids, or mating two single haploid yeast fusion plasmid transformants into a ... exacerbated by expression of target proteins in heterologous systems (e.g. mammalian proteins in S. cerevisiae...Addgene Browse Our Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About...activation. Vary the expression levels of bait and prey proteins - overexpression can lead to false interactions...During this period, proteins were successfully expressed as fusions that retain their individual activities...see Figure 2a) (4). These fusions are then co-expressed in a yeast strain that also contains a reporter...bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI’s interact, this...

  16. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    Published
    June 19, 2018, 1:38 p.m.
    ...Addgene.org Find plasmids for in vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent...Bourdeau et al created two constructs optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic... growth following acoustic collapse E. coli re-express arg1 gas vesicles and restore their ultrasound ...for ARG’s: in vivo imaging. Specifically, arg1 expressing E. coli aided in ultrasound imaging of the mouse...it’s a probiotic strain that can colonize the mammalian intestinal tract and has been used for 100 years... bowel conditions. In these experiments, arg1 expressing ECN were injected into the mouse colon, imaged...the images compared to mice injected with ECN expressing lux. arg1 and lux reporters both allow for visualization...

  17. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    Published
    Jan. 10, 2018, 4:13 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ...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...

  19. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...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,...
Showing: 21 - 40 of 162 results