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We narrowed to 162 results for: Mammalian expression plasmids

Showing: 141 - 160 of 162 results

  1. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...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....

  2. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...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. (...

  3. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...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 . ...

  4. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...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 ...

  5. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    .... 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...

  7. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...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...

  9. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...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 ...

  10. Typing CRISPR Systems

    Type
    Blog Post
    Published
    March 18, 2025, 1:15 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...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  ...

  12. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati, P., ...

  13. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ...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...

  15. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ... 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...

  16. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have... C. Transgene expression of transfected supercoiled plasmid DNA concatemers in mammalian cells. J Gene... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on... have been utilizing full plasmid sequencing services to verify the plasmids, including long-read sequencing... the sequencing results for their plasmids, they found the plasmids were much longer than expected, and...

  17. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ...Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains 1 kb homology...found within the homology areas of the plasmid to prevent the plasmid from being cut and destroyed by CRISPR... the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited...expensive for many labs. We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide...wide variety of genes with GFP. These plasmids are available through Addgene, and the stem cell lines we...found in the Cell Catalog on our data portal. The plasmids should be functional in many different human cell...constructs below. Read on to find out how these plasmids work and to discover more resources for your genomic...

  18. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...neuronal connectivity in the mammalian brain (Wickersham et al., 2007; plasmids at Addgene). AAV vectors...transgene expression. Fluorescent proteins are widely used for tracing and their expression is often made...Cre-dependent vectors are used in transgenic animals that express Cre in only specific cell types, for example only...labelling the subset of infected cells that also express Cre. Another advantage of AAV vectors is that there...et al. 2014 traced neurons with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection...Injection into a conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the...involved in learning and memory (Kohara et al., 2014; plasmids at Addgene). The Gradinaru Lab at the California...

  19. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...Differential cytosine editing between bacteria and mammalian cells Researchers in the Xueli Zhang lab were ...seemed to follow the same initial mechanism as in mammalian cells — by excising the freshly edited U bases...Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction. Nature...Additional resources on Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our...

  20. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...Developments in mammalian and yeast cells using modified Cas9 proteins to regulate gene expression (CRISPRi/...step as simple as heat-shocking to induce the expression of a Cre recombinase contained within the knock-in.... elegans. These modular toolkits build single plasmid targeting vectors that encode the gRNA transcript... C. elegans, visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to...
Showing: 141 - 160 of 162 results