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We narrowed to 237 results for: Mammalian expression plasmids

Showing: 101 - 120 of 237 results

  1. CRISPR/Cas9 FAQs Answered!

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    Blog Post
    ...high expression, so adding this G could help with expression, though it is possible for the plasmid to ...use double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase). The concept...easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer cloned ...you need to have four spacers co-expressed and using PX335 to express the Cas9n (nickase). Tips for using...articles (3/12/2014). With so many options for CRISPR plasmid tools and numerous experimental design decisions...concept of the double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase...backbone that uses the human U6 promoter to drive expression, is it necessary to add a G nucleotide to the...

  2. Fluorescent Proteins 101: Luciferases

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    Blog Post
    ...of light they emit or to optimize their expression in mammalian systems. Our luciferase collection page...677 61 AkaLumine ATP No Common uses for plasmids expressing luciferase Luciferase reporter assays for...gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression of luciferase...luciferase assays have become a gold standard in gene expression analysis to investigate the effect of regulatory...interactions Split luciferases, such as NanoBiT, are expressed in two subunits and only catalyze a luminescent...backbones with the luciferase gene, a variety of expression vectors, and literally thousands of reporter ... partnered with Promega to offer their Promega Plasmid Collection, including tools using NanoLuc®, NanoBiT...

  3. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    Blog Post
    ...and lentiviral expression vectors (Table 1). The four systems Name Function Plasmid Transduction...2019b). CHIME works through transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a...  The X-CHIME deposit also includes pXPR_071, a plasmid that leads to higher titer lentivirus preparations...first generation CHIME (pXPR_053). Find X-CHIME plasmids here!   Marty LaFleur is a Postdoctoral Fellow...Guide CRISPR: Protocol for Genome Deletions in Mammalian Cell Lines    ...

  4. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    Blog Post
    ...how your plasmid is intended to be used. Choose from: mammalian expression, bacterial expression, lentiviral...luciferase, cre/lox, yeast expression, worm expression, insect expression, plant expression, mouse targeting, ...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...

  5. Hot Plasmids and Viral Preps - January 2021

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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...control expression of their genes of interest. While recombinases are powerful gene expression tools in...changes in a protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...AAV-deliverable ABE.  Find the plasmid here Jesse Zalatan’s lab deposited plasmids for a chemically inducible...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...

  6. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    Blog Post
    ...high-copy plasmid-borne chromoproteins lost color overnight in liquid culture, though expressing the chromoproteins...for allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus...donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you’ve learned what chromoproteins...at the cells on the plate. For example, if your plasmid contains a gene encoding a blue chromoprotein, ...colonies to determine which cells have taken up the plasmid. Of course, other follow up methods like sequencing... chromoprotein sequences into E. coli BioBrick plasmids (meffRed, eforRed, asPink, spisPink, scOrange,...gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm...

  7. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...1053 amino acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency...capabilities of full-length SpCas9. This set of plasmids includes AAV-Cas9C-VPR for targeted gene activation...activation. Split inteins have also been used to express base editors. References Chew WL, Tabebordbar ...HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038...bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71 . https://doi.org/10.1038...In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9. Nat Biotechnol 33:102...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ...

  8. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

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    Blog Post
    ...neuronal expression. CAG-FLEX: CAG promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin...neuronal expression. hSynap-FLEX: Synapsin-1 promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP...to the PDGFR transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal...systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG...FLP-dependent expression GFAP: Glial fibrillary acidic protein (promoter). Good for Glial expression (not neurons...SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger fluorescent...dynamic range, photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants...

  9. The Challenges of Cell Culture

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    Blog Post
    ... Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about...Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ...along with Down-Regulation of HIF-1α by NDRG2 Expression Attenuates Proliferation and Invasion in Renal...

  10. Spurred by COVID-19, Addgene Bridges Plasmid Sharing Between Academia and Industry

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    Blog Post
    ...tagged and untagged expression of SARS-CoV-2 proteins in bacteria, yeast, and mammalian cells. We teamed ...Ginkgo Bioworks COVID-19 plasmid collection Beyond COVID-19: Addgene’s plasmid collections available ...COVID-19 plasmids to industry scientists in the short term, did you know we have other plasmid collections...organizations have made their plasmids available beyond academia. For example: Plasmids from the Michael J. Fox...and plasmids from several labs at Florida State University became openly available. Find plasmids available... more about the Ginkgo COVID-19 plasmid collection Read about plasmids in the COVID-19 collection ...one critical part was missing when the COVID-19 plasmids first became available: distribution was limited...

  11. Another Pathway into Cells: iTOP

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    Blog Post
    ... mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient...control could enter cultured cells and activate expression of a luciferase reporter. CPP was not required...stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR...causes a cell to take up, and continued protein expression would require multiple transduction events. For... to binary systems in which transient protein expression has a long-lasting effect, such as Cre- or Cas9...10.1016/j.cell.2015.03.028. PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters... Introduction of macromolecules into cultured mammalian cells by osmotic lysis of pinocytic vesicles. ...

  12. Microbes: Look, Listen, and Tell

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    Blog Post
    ...Resource Page CRISPR plasmids designed for use in bacteria Deposit Your Microbiology Plasmids with Addgene ... technology is widely applicable as bacterial expression of dyes allows dye visualization in vivo using... for noninvasive imaging of microorganisms in mammalian hosts." Nature 553.7686 (2018): 86.PubMed PMID...

  13. How to Deposit Your Plasmids with Addgene

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    Blog Post
    ...Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that will not be published are...and entering a plasmid name,plasmid type, and plasmid description/experimental purpose (right). On the...with your plasmids, and adding all the plasmids at once increases the likelihood that the plasmids are given... vector to be used in mammalian cells, check Lentiviral and Mammalian Expression). This information will...easy for you to share plasmids. To achieve this goal, we will archive any plasmids you've deposited with...Spreadsheet". Start your plasmid deposit There are two options to begin your plasmid deposit. Figure 2: Choose... Choose to submit plasmid(s) online or by using a spreadsheet. The plasmid information can be entered ...

  14. Hot Plasmids: Winter 2025

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    ... and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only for future...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...cell-derived neurons expressing ASAP5-Kv. C) Cultured rat hippocampal neuron expressing pan-membrane ASAP5... (Janicot and Maziarz et al., 2024). Each plasmid expresses all the components of the biosensor in optimal...selectivity. With 10 plasmids available as a kit, plus 21 optional accessory plasmids, the collection targets...precise plant breeding. Find dicot prime editing plasmids here! Vu, T. V., Nguyen, N. T., Kim, J., Song...Figure 2), the authors developed a cost-effective expression and labeling approach using these constructs ...

  15. Isolating Genomic Regions of Interest with the CRISPR System

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    ...biotinylation tag expressed on a plasmid, allowing for in vivo biotinylation through a plasmid-based biotin... The Fujii lab demonstrated this in mammalian cells by expressing dCas9 with an N-terminal 3xFLAG tag ...gRNA, which can easily be cloned into a gRNA expression plasmid. Experimental use Since the initial report...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...generated a transgenic mouse line and retroviral expression system for using this approach in in vivo studies...DNA-binding molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii lab also developed...imprinting (Zhang et al., 2017). This upregulates expression of IGF2, a characteristic of many human malignancies...

  16. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    ...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ...labeling, BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines... et al saw different levels of fusion protein expression and biotinylation depending on which terminus... requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein and saturation of... al allowed 24 hours for tetracycline induced expression and biotinylation of interacting proteins. This...Interested in using Split-BioID with your research? Plasmids are available from Addgene! References Schopp... Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann, J., Derua, R.,...

  17. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

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    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.        A retroviral...libraries have been created in lentiviral systems for mammalian screens, which enables researchers to maximize...type 2 (Th2) cells from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation...differentiation and activation, using expression of differentiation factors as the screen readout. This retroviral...treatment of cardiovascular disease. Find the LipoGlo plasmids at Addgene! Lawler et al, QJM: An International...

  18. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...Find tools for mammalian RNAi Browse all plasmids for RNAi Check out Addgene's plasmids and resources ...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence...mentioned above and is a popular RNAi trigger in the mammalian research community. Fortunately, many tools for...harvested instead of microscopic isolation. The plasmids required to conduct an APT-based RNAi experiment... target sequence, as well as essential control plasmids. Many thanks to our guest blogger Robert Orr...

  19. Viral Vectors 101: An Introduction to AAV

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    Blog Post
    ... to co-express them from one promoter. Specificity The ability of AAV to drive gene expression in specific...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression via switches...estimation. Once expressed, viral cargo is usually quite stable and in some mammalian tissues has shown...transgene expression, since you don’t have to wait for the host to replicate the DNA before expression. Now...non-enveloped icosahedral capsid, that can be used to express genes of interest in cell and animal models. AAVs...laboratory experiments ranging from transient gene expression in a particular cell type to CRISPR genome editing...genes, you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb. Co-infection rates...

  20. Giving gRNAs a Facelift - Synthetic and Beyond

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    ...are slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing...drawback of CRISPR-Cas9 technology is that most expression systems are constantly “on”. There is no mechanism...RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021)....ready-to-use viral preps of select CRISPR lentiviral plasmids Additional resources on the Addgene Blog  How...
Showing: 101 - 120 of 237 results