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We narrowed to 363 results for: nsf

Showing: 81 - 100 of 363 results

  1. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...genome edited plants via Agrobacterium-mediated transformation. We also constructed smaller vectors suitable...suitable for transient assays using protoplast transformation. Cytosine base editors for plant There are several...

  2. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...DNA-binding protein. The ADD domain of the methyltransferase binds to H3K4me0 on D3L, releasing the autoinhibition...

  3. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...certain modifications to DNA bases.  Ligate and transform your resultant plasmid libraries into chemically...chemically competent E. coli and analyze transformants for appropriately sized inserts. We consider a library...culture, dilute and plate the complete library of transformants for single colonies (this usually requires 10...10-20 plates) (see box below). This initial transformation should be performed using a highly competent...extract plasmid DNA directly from the colonies. Transform the resultant plasmid DNA into E. coli BL21 (DE3... protein production), then dilute and plate transformants for isolated colonies (150-200 per plate) across...

  4. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...we have found that transfecting with PBx mRNA can boost efficiency over transfecting with plasmids in some...from a licensed distributor) allow you to quickly transfer a DNA sequence from the transposon vector to one...traditional method of making stable cell lines is to transfect, select, and isolate clones. This procedure is...integration efficiencies that are accomplished with transfections and are therefore virus-free. This is achieved...fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur in your cargo...

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

    Type
    Blog Post
    Published
    June 23, 2020, 1:15 p.m.
    ...worked closely with the legal partners and Tech Transfer Offices at universities to make COVID-19 research-related...United States, and many universities and Technology Transfer Offices were, at least, temporarily closing, these...scientists, we’ve worked with many scientists, tech transfer offices, research foundations, and funding initiatives...them available to industry, please reach out at techtransfer@addgene.org and we will initiate a conversation...conversation with you and your technology transfer office to discuss the possibilities for expanded sharing. ...

  6. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...Interest (GOI), and transfect it into your favorite cell line. In parallel, you transfect cells with another...express your protein of interest endogenously and transfect them with a plasmid encoding your protein of interest...expect to see evidence of antibody binding in the transfected cells but not in control cells. But (there is...results show a lovely, strong band from the cells transfected with your protein of interest and show just the... the faintest band from the cells transfected with the other plasmid. Your antibody works! Multiple (Independent...

  7. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which...fluorophore. The difference for BRET is that the energy transfer is between a luminescent donor and a fluorescent...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha...transducers. To use the TRUPATH system, cells are transfected with plasmids expressing the GPCR of interest...

  8. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    Published
    July 22, 2025, 1:15 p.m.
    ...luminescent) worlds Forster Resonance Energy Transfer (FRET) is the transfer of energy from an excited donor fluorophore...Resonance Energy Transfer (BRET) works the same way, except that energy is transferred from a luminescence... fluorescent protein close enough, energy is transferred by BRET and the fluorescent protein emits light...

  9. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...strong enough to hold a plasmid together through transformation, so the cloned vector must be ligated. But ...nicks easily repairable by the bacteria after transformation. LIC is a reliable cloning method, but it is...recombination intermediate to bacteria, so it can be transformed as-is into cells. The cells then get to work ...is long enough to hold it together, it can be transformed into bacteria, which will efficiently repair ... annealed. The recombination intermediate is transformed into bacteria, which repair the plasmid. Created...

  10. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ... Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction... non-replicative. Nanoblades are produced by transfecting the following four components into 293T cells...lower off-target effects when compared to DNA transfection of gRNA and Cas9 expression plasmids. These ...having lower off-target edits compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades...can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote. As a proof-of-principle...

  11. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...easy — our tech transfer team will communicate with your organization’s technology transfer office or other... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...require that you send your samples to Addgene pre-transformed in this strain. Hazardous If the plasmid...samples to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new...

  12. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...demethylation?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental... al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun 9, 597. https://doi.org...promoters by targeted steric blockage of DNA methyltransferase with CRISPR/dCas9. Nat Commun 12, 5711. https...

  13. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...expression vector that is then transformed into a bacterial cell. Transformed cells propagate, are induced... strategy for protein expression consists of transfecting cells with your DNA template of choice and allowing...SV40, CMV, and RSV) for robust expression post-transfection. Mammalian systems can express proteins both... Both methods produce high protein yields if transfection is successful. Some mammalian systems also ...

  14. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...used by respondents (transfection method, amount of each component per transfection etc.). Take Pooran'...an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression...delivery methods. Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA...researchers will have to carry out repeated transfections with the ssDNA guide, although many may find...

  15. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...ccdB gene die off and a higher percentage of transformants contain the ccdB-less construct. Other toxin-antitoxin...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...specific nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...

  16. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...determined by the different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids...easily cloned in different bacteria, and can be transferred to various experimental hosts (Jain and Srivastava...scientific community, as they allow for simplified transformation processes, and can be used by scientists across...conjugative and carries the genes necessary for transfer initiation between bacterial hosts. Also, self-transmissible...Plasmids 101: What is a Plasmid?  Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Tranfection...

  17. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...size, multimers may exhibit reduced transformation and transfection efficiency compared to the monomeric...plasmid multimerization is to avoid growing the transformed bacterial culture into the late stationary phase...the linearized band, ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form... Hartl D, Rudolph C. Transgene expression of transfected supercoiled plasmid DNA concatemers in mammalian...

  18. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...proteins needed for the virus to replicate,  and the transfer plasmid which contains the viral genome. In today...Replication initiates from the AAV genome ITRs on the transfer plasmid, which contain regions of dsDNA. To generate...out this blog post to learn more about the AAV transfer plasmid! References: Naso MF, Tomkowicz B, ...was engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about...

  19. What the HEK?

    Type
    Blog Post
    Published
    April 18, 2023, 1:15 p.m.
    ...1970s by Alex van der Eb in the Netherlands. Transformation of HEK cells with adenovirus 5 DNA was postdoc...replication. To generate a virus, you need only transfect the ‘helper’ plasmids (viral packaging machinery...Protein production  HEKs are extremely easy to transfect in bulk and produce recombinant proteins at high...Resources References Shaw G., et al., Preferential transformation of human neuronal cells by human adenoviruses...

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

    Type
    Blog Post
    Published
    Oct. 29, 2015, 2:30 p.m.
    ...understand genetic cytoplasmic factors that could be transferred between cells. At the time, these extranuclear...-factors. Like F-factors, R-factors could be transferred between bacteria via cell-to-cell contact; however...pSC102, with EcoRI and selected for E. coli transformants that were resistant to both. When this proved...Chizuko Ogata, and Sachiko Sato. “EPISOME-MEDIATED TRANSFER OF DRUG RESISTANCE IN ENTEROBACTERIACEAE VIII:...
Showing: 81 - 100 of 363 results