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We narrowed to 237 results for: Mammalian expression plasmids

Showing: 81 - 100 of 237 results

  1. Inntags: Innovative Protein Epitope Tagging

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    Blog Post
    ...about other Protein Tags in our Plasmids 101 Series. Get the Inntag Plasmids Tag endogenous proteins with... in a scientist’s toolbox. As we’ve covered in Plasmids 101, tags can help you determine localization ...protein of interest, purify it, or determine its expression level without the need for a custom antibody....testing indicated that inntags IT5, IT6, and IT10 expressed well in cells and did not form undesirable aggregates... high correlation for protein localization in mammalian cells." Nature methods 10.4 (2013): 315-323. Pubmed... size or viability compared to untagged GFP overexpression, while the rest of the fusions had some negative...

  2. Cpf1: A New Tool for CRISPR Genome Editing

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    ...Cpf1 orthologs that display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...Cpf1 nucleases would exhibit in vivo activity in mammalian cells. From 16 diverse Cpf1 candidates, Zetsche...multiplexing approach using a single crRNA array to express up to 4 crRNAs. Zetsche et al. also suggest that...efficiency of two Cpf1 orthologs known to be active in mammalian cells, LbCpf1 and AsCpf1. In both reports, on ...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...

  3. Google Forums Round Up: First Impressions of NgAgo

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    Blog Post
    ...cells with 5’-P-ssDNA guides and an NgAgo expression plasmid to edit a gene in vivo. Try NgAgo for Yourself...gRNA complexes or with guide RNA- and Cas9-expression plasmids. Cas9 guide RNA delivery does however rely...sequencing on select regions of all distributed plasmids as part of quality control, but does not perform...unlike an RNA guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo... dangerous, non-physiological temperature for mammalian cells and one which diminishes the endonuclease...(1), NgAgo has received much attention and the plasmid has been requested by labs around the globe nearly...can indeed be optimized for genome editing in mammalian cells. However, the fact that most of the researchers...

  4. Plasmids 101: Control Plasmids

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    Blog Post
    ...Knockdown the expression of Gene X using shRNA expressed from a plasmid   Figure 1: Expression Level of Gene... used for plasmid-based experiments. What are control plasmids? Generally, control plasmids help to ensure...as the expression level of Gene X in cells treated with Plasmid A is similar to the expression level of...follow an example of a plasmid-based experiment, delineate its requisite control plasmids, and discuss why ...proper control plasmids would address and answer these questions. Types of control plasmids Part of planning...efficiency. An internal control may be a plasmid that constitutively expresses a reporter protein (e.g., GFP or...Figure 1, an internal control, such as the GFP-expressing Plasmid B, could demonstrate whether the cells were...

  5. Plasmids 101: Inducible Promoters

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    Blog Post
    ...Resources on the Addgene Blog: Plasmids 101: The Promoter Region Plasmids 101: Cre-lox Light-Switchable...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...lac inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is ...Promoter Subtype Promoter Example Activator Addgene Plasmid/Kit Alcohol-regulated AlcA promoter AlcR pGGA008...Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than chemically...chemically induced promoters; they show near-zero expression at regular temperatures but can be induced by ... promoters, in which a gene of choice is only expressed following exposure to a brief heat shock. In the...

  6. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ...because they can be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs...scFvs in mammalian, yeast, plant, and insect cells, they are most often expressed from a plasmid in bacteria...Ahmed et al., 2012). One challenge of bacterial expression is the proper formation of the disulfide bond...requires an oxidizing environment to form. When expressed in the non-oxidizing cytoplasm of bacteria, scFvs...more of these approaches are often used for scFv expression in bacteria:  targeting to the oxidizing periplasm...to allow for proper disulfide bond formation.  expression in strains of bacteria that are redox mutants...mutants so they have a more oxidizing cytoplasm. expression in the presence of molecular chaperones that help...

  7. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...permanently induce expression? Check out our Retro expression vector and Lenti expression vector. Turn it...resort to the force to control protein expression in mammalian cells: easy to use biological tools are...ablate protein expression. If you need the ability to control when you turn off expression for good, the...inducible expression (more on this below) will both achieve temporary repression of protein expression. Addgene... a gene (cDNA) can be done by either plasmid or virus. A plasmid carrying your cDNA can be targeted for...appropriate promoter to express your cDNA, depending on the level of expression you would like. Check out.... Whether you want to express or repress, good luck on your protein expression journey! Resources and...

  8. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

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    Blog Post
    ... resources on the Addgene blog Plasmid 101 Protein Expression Plasmid 101 Biotinylation Other resources...proteins per cubic micron in bacteria, yeast, and mammalian cells (Milo, 2013). The number of interactions...capture protein complexes, scientists can simply express a bait protein fused to BirA* in cells and then...vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development of BioID and... specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate peroxidase...sensitivity. Sensitivity could be increased with higher expression levels but for many fusion proteins this would... They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid and non-toxic proximity...

  9. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    ... about FLEx vectors Download our Plasmids 101 eBook Read our plasmids blog posts Resources on the Addgene...molecular biology reference Find Cre-lox plasmids at Addgene Browse plasmids at Addgene ...optical and genetic methods to photo-stimulate mammalian neurons and alter synaptic events: a new technology...not naturally light-sensitive, so the selective expression of opsin genes in targeted cells makes it possible...How to get an opsin into the brain In order to express opsins within the brain, researchers inject genetically...relatively stable extra-chromosomally long-term expression and low ability to induce innate immune responses... promoters; however, those usually drive weak expression of downstream genes. Strong cell-specific promoters...

  10. Hot Plasmids - May 2022

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    Blog Post
    ...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...protein can then be co-expressed in essentially any transgenic zebrafish line expressing a GFP-tagged version...display increased neuronal expression in the brain while having decreased expression in the liver after systemic...only AAV.CAP-B10 causes brain wide expression, with no liver expression, after retro-orbital sinus vein ...need. If you'd ever like to write about a recent plasmid deposit please sign up here. Here's what you'...in vivo proximity labeling Increased neuronal expression using Cre-mediated AAV Targeted Evolution method...

  11. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

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    Blog Post
    ...cloning, and plasmid tags, addtional content added to the "Plasmids that Glow" and "Viral Expression Vectors...tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI...different bacterial (and eventually mammalian) species were cloned into plasmids and new cloning vectors such...topology, and cloning and gene expression. Just released: Addgene's Plasmids 101 (2nd Ed.) eBook And now...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...factors played within an organism.  So how did plasmids get their name? In 1952, Joshua Lederberg set...inheritance factors. He proposed the catch-all term “plasmid” derived as a hybrid of "cytoplasm" and "id" (Latin...

  12. What the HEK?

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    Blog Post
    ...because they express the SV40 large T-antigen. This allows them to replicate viral plasmids with the SV40...transfect the ‘helper’ plasmids (viral packaging machinery) along with the viral plasmid containing the cargo...derivatives of HEK-293, the most commonly used line expresses SV40 large T antigen and is thus dubbed HEK-293T...receptors and ion channels are often exogenously expressed and studied in HEKs due to the cell line’s ability...produce these proteins and its relatively low expression of native channels.  No matter what your research...factors that could skew research results (gene expression, growth rates, etc.). Thus, it’s important to...blog Five Popular Model Organisms 10 Tips for Mammalian Cell Culture Adenoviral Vector Production and...

  13. Plasmids 101: Using Transposons in the Lab

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    ...move antibiotic resistance genes between plasmids or from plasmids to the bacteria’s genome. In humans, if...Non-autonomous transposons, however, need another TE to express a transposase or reverse transcriptase so they ... has a close-to-random integration profile in mammalian genomes. SB is active in vertebrates and integrates...activity (Voigt et al., 2016). Sleeping Beauty plasmids  piggyBac Although its name suggests otherwise... well as in vivo in yeast, plant, insect, and mammalian cells, including human cells. piggyBac is biased...transposase (hyPB) is ~10-fold more active in mammalian cells compared to the codon-optimized wild-type...wild-type piggyBac transposase.  piggyBac plasmids    Tol2  Tol2 was the first reported active DNA transposon...

  14. Hot Plasmids and Viral Preps - September 2021

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    ...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...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available... Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter... be seen through the success of the change in expression of GFP, for the stop codon should have changed...

  15. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    Blog Post
    ... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain...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 the Hot Plasmids podcast ...pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing mice, and used ...responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed... genes for noninvasive imaging of microbes in mammalian hosts Article contributed by Lukas Morgan  Listen...

  16. 22 Hot Plasmid Technologies from 2014

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    ...Cre-based Brainbow-expressing cells. Autobow plasmids are all-in-one versions which express self-excising ... host cells in order to optimize expression. The expression plasmids for a given species are based on ... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...tested is expressed along with Cas9. If the gRNA effectively cuts the target sequence, the plasmid undergoes...designed the pOSIP (one-step integration plasmid) series of plasmids, vectors that convey both the sequence...Chen, pCS2-3xFLAG-NLS-SpCas9-NLS is a Cas9 expression plasmid that was used by Chen and colleagues for ...

  17. Plasmids 101: Cre-lox

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    ...previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters...codon is excised, and gene expression proceeds. One popular lox-stop-lox plasmid is from Tyler Jack's lab...Resources on Addgene.org Find Cre plasmids Find loxP plasmids  ...common uses are listed below. Cre-dependent gene expression – placing a series of transcriptional and translational...upstream of a gene of interest will prevent gene expression in the absence of Cre. In the presence of Cre...floxing”, for “flanked by loxP”), will permit gene expression until Cre is present, at which time the gene ...to easily perform this eviction. Regulated Cre expression – placing Cre downstream of promoters that are...

  18. Educational Planarian Tools for Teaching Developmental Biology Techniques

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    Blog Post
    ...RNAi) to knockdown expression of specific gene expression. The 49 deposited plasmids enable students to...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...protocol resources. Find Sánchez lab Plasmids Visualizing gene expression via Whole Mount In Situ Hybridization...insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new labs to...are transformed with the appropriate plasmid and dsRNA expression is activated through the  induction ...Ingestion of bacterially expressed double-stranded RNA inhibits gene expression in planarians." Proceedings...

  19. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

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    ...control RFP expression plasmid.  With this set up, all successfully transfected cells will express RFP while...using the supplied donor plasmid will express GFP. If there’s GFP expression, presumably the endogenous...H2B (histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter...and to make the desired genome edit:  A donor plasmid that contains a puromycin resistance gene linked...the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The Madison lab also created...inducible SRIRACCHA system (iSRIRACCHA) where Cas9 expression is activated using 4-OH-T and doxycycline. A ...repaired by NHEJ as well. The ratio of RFP to GFP expressing cells provides a real-time read out of Cas9 activity...

  20. Plasmid Tools for Microbiome Studies

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    ...can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be identified...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they... tools for microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to...
Showing: 81 - 100 of 237 results