Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 237 results for: Mammalian expression plasmids

Showing: 141 - 160 of 237 results

  1. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...of these FPs are fluorescent when ectopically expressed in most organisms. Furthermore, fusing FPs to ... The two most common uses are: 1) to test the expression level in a specific system (by measuring the ...optimized for 37°C, and is therefore most suited for mammalian or bacteria studies, whereas GFPS65T is better... parameter is important if you are planning to express the FP in acidic environments (e.g. yeast cytosol...He is interested in everything related to gene expression, particularly at the RNA level. He maintains ... comprehensive data sheets on many FP variants Plasmids 101: Green Fluorescent Protein (GFP) Three Review...proteins and imaging technologies to study gene expression, nuclear localization, and dynamics” Curr. Opin...

  2. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ... as a donor plasmid containing the transposon, along with a helper plasmid that expresses the transposase...lingering expression of reprogramming factors. Non-replicating Plasmids: Generating iPSCs with plasmid-based...plasmid-based expression requires serial transfection of 1 or 2 plasmids that express the reprogramming factors...reprogramming. Additionally, lingering expression or re-expression of viral transgenes in iPSC-derived cells... generating iPSCs by plasmid transfection where 2 of 11 clones tested had plasmid integration. Another...the dose of plasmid the cells receive over the whole reprogramming period and the plasmid will get diluted...larger plasmids have lower rates of transfection. Minicircles: Minicircles are like mini-plasmids. They...

  3. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...the ability to distribute plasmids with the piggyBac™ transposon. These plasmids, when combined with a source... minimize expression levels of your gene of interest, as needed. Controlling the expression levels of ...useful resource. While Addgene cannot distribute plasmids with the piggyBac™ transposase itself, please ...successfully used for genetic manipulation in mammalian cells until 2005 [1]. Like other transposons, ...genome fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur...may be particularly useful when the gene being expressed has threshold effects, such as with signaling ... can also mitigate the deleterious effects of expression of certain genes, such as those that have negative...

  4. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ... Visit Our Stem Cell Page Find Plasmids for Reprogramming Find Plasmids for Differentiation ...the embryonic and extraembryonic cell types. In mammalian development, cells found in the zygote through...reprogramming differentiated adult cells by the ectopic expression of four embryonic genes- OCT4, SOX2, KLF4, and...to a Nobel Prize for Dr. Yamanaka in 2012. Find plasmids for generating iPSCs iPSCs and ESCs closely resemble...possess similar morphologies, growth properties, and express stem cell markers similar to ESCs [11].  While ...

  5. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...native expression levels. However, it is advisable to first express the fusion protein from a plasmid-based...instance, Lifeact-mTurquoise2 (Plasmid #36201) or mScarlet-tubulin (Plasmid #85045). We often use restriction...fluorescent proteins that behave as monomers in mammalian cells include: mTurquoise2, mEGFP, mNeonGreen,...strategy (Sheridan, 2002). Avoiding protein over-expression artifacts The introduction of DNA that encodes...of protein within a cell and may lead to over-expression artifacts. This is an important downside of the...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...

  6. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...Browse fluorescent protein plasmids and resources See all Jaffrey lab plasmids ... cellular processes by transforming cells with plasmids encoding aptamer-tagged RNA and later introducing...aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...aptamers for imaging small non-coding RNAs in mammalian cells." Nature communications 9.1 (2018): 656....

  7. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...instead get expression of the sensor, marker, or whatever other tool you chose to express. In the case...most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses...that the virus must have its structural proteins expressed from a reverse-transcribed piece of DNA called...is done, and the transgenic genome will simply express from whatever promoters were included in the vector...And there you have it, the full “life cycle” of expression of selfish, lazy genes of a non-living viral ...Addgene's Lentiviral Guide Most popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (...

  8. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...2018). cDNA libraries, which consist of plasmids that over-express coding sequences from a given cell type...used in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes...: once the environmental trigger leaves, gene expression is no longer activated. However, there are several...up-regulated. For instance, generally, the more highly expressed a gene is under native conditions, the less activation... for altering DNA sequence or regulating gene expression.     References Behan FM, Iorio F, Picco G, Gonçalves...Tsherniak A, Kadoch C (2018) Interrogation of Mammalian Protein Complex Structure, Function, and Membership...Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell 159:635–646 . ...

  9. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...in the medium or flavin containing proteins in mammalian cells. Generally, autofluorescence is less of .... "A genetically encoded fluorescent probe in mammalian cells." Journal of the American Chemical Society... "Live cell imaging of phosphoinositides with expressed inositide binding protein domains." Methods 46.3...protein fusions Browse the Fluorescent Proteins Plasmid Kits ...

  10. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...activity. To drive selective A → T editing, they co-expressed Polη, a translesion synthesis polymerase known...precise, efficient A•T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4)...editors for efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547...reporters Resources on Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our...

  11. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    .... citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...knockdown expression of multiple target transcripts simultaneously. Marraffini lab's CRISPRi/a plasmids In addition...targeted repression of gene expression. They successfully knock down expression of essential and non essential...engineering functions in bacteria, but have no fear - plasmids from Addgene depositors are making it easier than... system available from Addgene consists of two plasmids: pCas9: carries Cas9 and chloramphenicol resistance...) technology (Ronda et al., 2016). pMA7CR_2.0 expresses lambda Red and Cas9, which are separately inducible...gRNA and a backbone-targeting gRNA cassette for plasmid curing through "self-destruction" after induction...

  12. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...Allen Institute for Cell Science Plasmid Collection Check Out Plasmids for FRET Check Out the Fluorescent...into a vector encoding a fluorescent tag. Once expressed as a fluorescent protein fusion, it is possible... Figure 1 for commonly used markers for major mammalian organelles and transport pathways. It is best,... on the gene names in the figure below to find plasmids containing that particular gene. ... For subcellular markers in yeast see our plasmid collection from Sue Jaspersen (Stowers Institute...Prescott (Monash University). You can find more plasmids for labeling your subcellular structure of interest...interest in the Allen Institute for Cell Science Plasmid Collection. You can also use the library of full-length...

  13. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...control for many of the effects of gRNA and Cas9 expression, they fail to account for the effects of Cas9...lentiviral vector to cell lines that either stably expressed Cas9 or that had been lentiviral infected with...sites should control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting.... 5. Bassik, Michael C., et al. "A systematic mammalian genetic interaction map reveals pathways underlying...on Addgene.org Check out this guide to pooled plasmid libraries Find all of the CRISPR Pooled Libraries...

  14. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...bacterial genome, and inserted it into E. coli to express and then test for any proteins that could cut RNA..., when they looked at the transformed E. coli expressing Cas7-11, they found no evidence of growth inhibition...similar to shRNA and Cas13 systems in a range of mammalian cell lines. However, in those same comparisons...Use PASTE can be delivered via a single dose of plasmids, works in non-dividing and primary cells, and ...

  15. Typing CRISPR Systems

    Type
    Blog Post
    ... you need to ensure every unit of the complex expresses at similar levels. It was only recently that researchers... researchers began building optimal plasmids to harness the power of Class 1 systems. Class 1 CRISPR systems...effector and is able to target and edit RNA in mammalian cells. Type IV Type IV is the middle child (like...Lastly, type IV systems are most often found on plasmids. The many unique features of IV-A and IV-B have...led to the hypotheses that they are involved in plasmid competition or can hijack machinery from other .... Cas9 was the first to be adapted for use in mammalian systems, with the landmark publication in 2013...highly diverse and involved in competition between plasmids. Nucleic Acids Research, 48(4), 2000–2012. https...

  16. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...as a mechanism of transient knockdown of gene expression. This system could be used in lieu of siRNA or...RNA cleavage and is significantly less toxic to mammalian cells than Cas13 (likely due to the lack of collateral...More resources on the Addgene Website CRISPR Plasmids and Resources CRISPR Guide  ...

  17. Common Injection Routes in Mice

    Type
    Blog Post
    ...Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati, P., ...

  18. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...Addgene.org Find the Spinach Plasmids at Addgene Find Broccoli Plasmids at Addgene View All Fluorescent...Broccoli cassettes, thereby extending their use into mammalian cells. They’ve also dimerized Broccoli to further...processing using broccoli reveals optimal aptamer expression strategies.” Chem Biol. 2015 May 21;22(5):649...

  19. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian Expression ...editing. How to use Addgene plasmids for Easi-CRISPR You can use Addgene plasmids to create a repair template...of Addgene plasmid options for templates - see the example at right using blue flame plasmid pmScarlet_...desired insert (e.g., mScarlet).   Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N ...pre-CRISPR was very arduous. First, you’d construct a plasmid with your desired changes, long homology arms and...template! If you have a plasmid with your desired insertion feature (e.g., a fluorescent protein), you can...described in Figure 2.  You can also use Addgene plasmids to purify Cas9 protein. The table below contains...

  20. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ... have proved challenging to study. Cells often express multiple integrins that bind the same ligand, making... conformational complexity There are 24 known mammalian integrins, all heterodimers composed of one of...combination, showing that among the three αV integrins expressed on the BJ-5ta fibroblast cells, integrin αVβ1 ... readily accessible recombinant antibodies and plasmids. To encourage collaboration and education in protein...
Showing: 141 - 160 of 237 results