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We narrowed to 522 results for: des.2

Showing: 241 - 260 of 522 results

  1. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment, they found the ... cells throughout the body, especially stem cells 2. Remove any immunogenicity of the AAV vector Once ...Science. 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H...dystrophin transcript. Unfortunately, the oligonucleotides only modestly improve muscle function, and they must...wasting leading to death at around 30 years of age. Despite the amount of research conducted on DMD, there ... over time, stem cell editing would be highly desirable. Should stem cell editing occur, the CRISPR components... for Duchenne muscular dystrophy. gRNAs can be designed to flank a single or multiple exons that, when...

  2. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...notebooks you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn...with NEB enzymes, the app is quite helpful and provides a list of directions including which buffers to...

  3. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ...immunization until the desired level of antibodies is reached - typically within 2-4 months. After the animal...animal is given additional booster immunizations every 2-3 weeks to increase the antibody titer. The titer ... animal has the desired level of antibodies in its bloodstream, the antibodies are retrieved first by ...species. Pre-adsorbed polyclonal antibodies are designed for experiments where antibodies raised in several...

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

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    .... pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while pCRISPomyces-2 contains Cas9 and...pCRISPomyces-2 displays a higher editing efficiency, perhaps due to its condensed design. For both systems...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...Marraffini’s lab took advantage of this lethality to design the first synthetic bacterial CRISPR system in ...gRNA cassette for plasmid curing through "self-destruction" after induction with L-rhamnose and aTet. CRMAGE...scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering with ...

  5. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    Published
    Feb. 2, 2021, 2:15 p.m.
    ...and scholarships in different subject areas!    2. Resume and cover letter know-how  Tailor your resume...4. Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch... such as citizenship status, academic year, and grades that serve as automated filters of consideration...and cover letter revised by your network, which includes your friends, family members, and professional...

  6. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ...per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, R6K, and ColE2 plasmids) as cell division...biotechnology 36.4 (2018): 352. PubMed PMID: 29553576. 2. Kittleson, Joshua T., Sherine Cheung, and JChristopher...of copy number (Segall-Shapiro et al., 2018). Designing stabilized promoters To keep gene expression stable...ideally be optimized to promote generation of the desired product while avoiding the accumulation of toxic...

  7. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...without leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing...with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D...facilitate the selection of cells containing your desired edit. The selection is marker is then seamlessly...sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,”...Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted and unwanted target locus modifications...

  8. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...linear fashion. This implies that reducing the power 2-fold does not reduce photobleaching by half. How exactly...prolonged time). Other illumination strategies, such as 2-photon-excitation, selective plane illumination, TIRF...different ways. We describe a number of issues with some of the experimental designs used to make these...of Amsterdam. The previous post in this series described a practical approach to selecting a bright fluorescent... protein. Photobleaching is the irreversible destruction of a fluorophore under the influence of light...photobleach at some point. For live-cell imaging, it is desirable to have fluorescent proteins that are photostable...photoswitching (Kremers et al, 2009), which usually are undesired properties. In the ideal situation, a fluorescent...

  9. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    Published
    Feb. 11, 2021, 2:15 p.m.
    ...plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 gamma subunits of the heterotrimeric G...activation via bioluminescence resonance energy transfer 2 (BRET2). Check out our blog post on GPCR signaling...new receptors were termed Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) (Roth, 2016...What do smell, taste and sight have in common, besides being one of the five senses? GPCRs or G-protein-coupled...photons or odorants, to elicit downstream signaling cascades.  So let’s break down the name "G-protein coupled...then induce or inhibit intracellular signaling cascades as a response to the extracellular stimuli. Ligand...

  10. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    Published
    Oct. 21, 2025, 1:15 p.m.
    ... engineered components, like protein tags (Figure 2). Through the work of many groups, there are systems... in Addgene's repository (Table 1). Figure 2: Adding function to My Favorite Protein (MFP) through...protein innovations for therapeutic and vaccine design. We all want tools that expand what we can achieve...features with optimal biological performance are described as Click Biology (Howarth, 2025). Figure...nucleic acid sequence (Addgene can help with this!). Besides requiring stable covalent reactions, other required... Addgene. Addgene Link Click Biology Tag Description Humberg et al. (2025) Split inteins Cysteine-less...  Click Into Biological Building Click Biology describes the search for robust, reliable reactions, using...

  11. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...residues, which can be found throughout the antibody. 2) Endogenous cysteines, such as those that make up ...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 ...we would be remiss not to at least mention. Oligonucleotides allow for sensitive detection of low abundance...strong protein-protein interactions with other peptides, like an Fc-binding domain (Figure 1).  ...Strong protein-protein interactions with other peptides. Created with BioRender.com.     Many biologists... conjugation. But hopefully, this introduction provides a good base from which to dive deeper!References...

  12. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ...successfully regulated expression of channelrhodopsin-2, commonly used in optogenetics, opening up the possibility... Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation...GFP-labeled cells. Once Tang and Cepko found a description of GFP-binding nanobodies, the project really...stable inside cells. Nanobodies binding GFP were designed in 2009. Subsequent work showed that nanobodies...green-red Cre-lox systems. T-DDOGs can also be designed with other DNA-binding domains, including the ...

  13. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...of R bodies R bodies have been studied for many decades, and they were known to be resistant to detergents...

  14. The Twelve Days of CRISPR

    Type
    Blog Post
    Published
    Dec. 22, 2020, 2:15 p.m.
    ...Addgene gave to me: five methods for detecting SARS-CoV-2 using CRISPR.  On the sixth day of CRISPR, Addgene...#12DaysOfCRIPSR hashtag to share new CRISPR content designed to help you with your CRISPR experiments and to...CRISPR, Addgene gave to me: tips and tricks for designing gRNAs for CRISPR editing. On the fourth day of...

  15. New Videos: Addgene Lab Tips

    Type
    Blog Post
    Published
    Feb. 15, 2018, 2:41 p.m.
    ...this quick trick. Just flip, and press! Lab tip #2: Tube flick trick When combining reagents in the lab...distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep track of reagents in your...materials quickly and easily. Learn more about using barcodes. Check out our “Lab Tip” filter on the blog...

  16. Twenty Years of Sharing Science!

    Type
    Blog Post
    Published
    Jan. 8, 2024, 3:25 p.m.
    ...anniversary! We've done so much in the past two (2!) decades that it's hard to summarize, but we wanted to...experience we're sharing through blogs, eBooks, videos, guides, protocols, and of course, the friendly folks at...

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

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...in one direction. Cas3 is thus not helpful for designer genome edits, but it is effective at wiping out...David, Chen, S., Janice, et. al. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Science, 362...

  18. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ... follow the same general cyclical process (Figure 2). The differences typically come in when generating...starting point for the next cycle. Figure 2: General process of antibody directed evolution. Created...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute... antibody sequences, degenerate primers can be designed to selectively mutate specific regions of your...sections. Structure-guided directed evolution If you desire a bit more control, structure guided directed evolution...complex as a template. This structural framework then guides the specific mutations you choose, whether it's...allows you to combine the strengths of rational design with the power of directed evolution, all while...

  19. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...somewhere else (Fig. 2). Class II TEs have LTRs on both of their ends.    Figure 2: Overview of DNA... four major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8, Cas7, Cas6) that forms ... sequence that moves around. This DNA sequence encodes the proteins the transposon needs for transposition...contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase and integrase enzymes...ORFs) which often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities...preferred target site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from...encode reporter genes, mutagenic cassettes, or barcodes. When transposons are delivered to cells or a ...

  20. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...an internal promoter, and the desired gene (transfer vector plasmid), 2) the HIV-1 genome containing all... several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one that ...PubMed PMID: 3785217. PubMed Central PMCID: PMC367857. 2. Naldini, Luigi, et al. "In vivo gene delivery and...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging cell lines to avoid recombination...
Showing: 241 - 260 of 522 results