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We narrowed to 237 results for: Mammalian expression plasmids

Showing: 21 - 40 of 237 results

  1. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org Check... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected... mammalian cells...

  2. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ...reconstituted the EGFP expression cassette. (b) pCAG-EGxxFP target plasmid and pX330-sgRNA plasmid (Mashiko et al...fragments. The pX330 plasmid contains humanized Cas9 expression cassette and sgRNA expression cassette. The ...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ... mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA." Scientific reports...to Help Design You gRNA Find CRISPR Plasmids for Use in Mammalian Cells Check out All of Our CRISPR Resources... has expanded since its adaptation for use in mammalian cells in 2012-2013. Researchers are now using ...

  3. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...human gene for expression in E. coli. Genes are also optimized for expression in mammalian cells. For example... for expression in mammalian cells. This was the first codon optimized membrane protein expressed in mammalian... usage bias in Plasmids 101: Codon usage bias Read more about plasmids in our Plasmids 101 blog series...Cells however, may not express all 61 of these tRNAs and those tRNAs that are expressed may be found at very...extremely useful in expressing functional proteins in hosts that do not naturally express that gene. The protein... mammalian cells. In this study, codon optimization resulted in a 6-9 fold increase in expression of these...GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal of Neuroscience...

  4. CRISPR 101: Epigenetics and Editing the Epigenome

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    Blog Post
    ...loci. Charles Gersbach’s lab has deposited mammalian expression constructs including pcDNA-dCas9-p300 Core... targeted cytosine demethylation in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to...for targeted cytosine methylation in mammalian cells. Co-expression markers EGFP and PuroR enable sorting...vector in chick embryos, it also functions in mammalian expression systems.   Christopher Newgard’s Lab has... are an additional layer of control over gene expression that go beyond genomic sequence. Dysregulation...modifications are correlated with alterations in gene expression, but causality and mechanisms remain unclear....that help package DNA, were associated with gene expression or silencing. To alter DNA modifications, researchers...

  5. Hot Plasmids - February 2022

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    Blog Post
    ...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'...

  6. MXS Chaining

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    Blog Post
    ...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...

  7. Plasmids 101: Terminators and PolyA signals

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    Blog Post
    ...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...

  8. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...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 ...

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

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    Blog Post
    .... 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...

  10. 15 Hot Plasmids from 2017

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    Blog Post
    ...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...

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

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    Blog Post
    ...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...

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

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    Blog Post
    ...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...

  13. Antibodies 101: Producing Recombinant Antibodies

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    Blog Post
    ...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...

  14. CRISPR 101: Multiplex Expression of gRNAs

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    Blog Post
    ...  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...

  15. 10 Basic tips for mammalian cell culture

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    Blog Post
    ...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...

  16. Antibodies 101: Affinity Tags

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    Blog Post
    ... 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...

  17. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ... 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...

  18. What's New in CRISPR - November 2021

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    Blog Post
    ...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...

  19. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...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...

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

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    ...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...
Showing: 21 - 40 of 237 results