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We narrowed to 889 results for: ANT;

Showing: 301 - 320 of 889 results

  1. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ... CRISPR’s therapeutic potential in mice with a dominant cataract disorder caused by a single copy of a...co-injection of Cas9 and a single-guide RNA targeting the mutant allele. When all was said and done, the researchers...therapies that are currently approved or have made significant progress in clinical trials.     Figure...almost all patients experiencing sustained and substantial relief from both disorders. As of publication...effectiveness, and delivery. Patient safety is important for allogenic therapies to prevent a graft vs....vs. host response. Off-target effects are also important safety considerations for all CRISPR therapies...Thus far, several targeted therapies have taken advantage of what our current delivery technologies are ...

  2. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...delivered using viral vectors! One of the most significant advantages of viral delivery is the potential for ...Calcium is arguably one of the most important metal ions in biology (sodium and potassium are up there...start by asking ‘What is the biological phenomena I want to observe?’ This information, along with a little...your experiment could be subtle or dramatic. You want to use a calcium sensor that is most sensitive to... than in the cytosol. Thus, for the ER you would want a low affinity calcium biosensor, so that the (1... with an extra apoCaM-binding motif to prevent aberrant binding of the biosensor to endogenous calmodulin...CEPIA) or through FRET sensors, can reduce these unwanted variables. Twitch is a FRET-based calcium reporter...

  3. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...Taking advantage of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes an elegant system...also be applied to other fluorophore/antibody pairs. Because the anti-GFP nanobody cross-reacts with BFP...length can and should be varied as it can have important implications for the fusion protein effectivity...GFP on the surface and “receiver” cells express an anti-GFP nanobody on the cell surface. When the cells...both spatiotemporal protein degradation and quantification of this depletion using imaging-based measurements... the degradation of the AID-tagged protein. To quantify protein degradation the Ward lab used fluorescent...levels of dCas9 expression and contain different antibiotic resistance markers. New CRISPR pooled library...

  4. Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana

    Type
    Blog Post
    Published
    Dec. 20, 2016, 3:30 p.m.
    ... as a successful applicant to the Addgene, Seeding Labs Plasmid Grant. For this grant, we’re working with...about the Addgene, Seeding Labs Plasmid Grant Read about Plasmid Grant Recipient, Gladys Kaba Resources on...their research. Kwabena came across the plasmid grant in Seeding Lab’s Newsletter and recently visited...Kwabena applied for the Addgene, Seeding Labs plasmid grant. 7:12 - 7:11 : Kwabena's work in Christopher E. ... recipient of the Addgene, Seeding Labs Plasmid grant and is working on a variety of projects in Ghana...

  5. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...Technique Sequence Information Detection Limit Advantages Disadvantages Mismatch Cleavage Assay Not Provided... Technique Detection Limit Application Advantages Disadvantages GUIDE-Seq 0.1% Cell-based  Searches ... the University of Cambridge. One of the most important steps in the CRISPR experimental process is validating...silico prediction. These techniques differ in important ways, but can also complement one another by providing... amplicon sequencing (Hendel et al. 2015), a significant difference as CRISPR experiments move closer ...networks in biological systems. NGS will also play significantly into the realm of clinical therapeutic development...sequencing: a comparison of methodologies for KRAS variant detection." Biotechniques 53.3 (2012): 182-188....

  6. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...guest bloggers, Jae Lee and Pantelis Tsoulfas. Jae K. Lee is an assistant Professor at the University ...This post was contributed by Jae Lee and Pantelis Tsoulfas of the Department of Neurological Surgery ...arrows point the direction of the specimen, A= anterior, P= posterior, D= dorsal, V= ventral Developments... milliseconds to acquire one image, thereby significantly reducing the time it takes to image through ...Similar to light microscopy, there have been significant advances in tissue clearing techniques during...which is a hydrogel-based method that also allows antibody penetration for immunohistochemical labeling in...Medicine. Find plasmids from the Lee Lab here. Pantelis Tsoulfas is an Associate Professor at the University...

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

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ... using a dCas9 variant as well. Furthermore, they provide a high-fidelity (HF) variant of the BE3 that...Addgene depositor Luciano Marraffini’s lab took advantage of this lethality to design the first synthetic...the original pCas/pTargetF but has many other advantages including shorter plasmid curing time. pCRISPomyces... a mammalian cell line (Qi et al., 2013). Two important features of this early system are its regulatable...bacteria. The authors were able to engineer a dCas9 variant fused to bacterial enhancer binding protein required...could be genes, specific polymorphisms, including antibiotic resistance genes. This approach successfully ...Streptomyces Some of the most industrially and clinically relevant bacterial species reside in the genus Streptomyces...

  8. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    Published
    Dec. 29, 2020, 2:15 p.m.
    ...for AAV Quantitation by Kate Harten DeMaio We made the switch to droplet digital PCR to quantify AAV titers...development virtually. 6. Quantifying DNA? Here are Five DNA Quantification Methods to Consider by Jennifer... dominates DNA quantification discussions, but what about the other methods to quantify DNA? Here we cover...Methods Based on CRISPR/Cas by Shravanti Suresh Guest blogger Shravanti Suresh who studies CRISPR at Iowa...

  9. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...challenge when I started working with AAV vectors. I wanted to use an AAV vector to express ChR2(H134R) in ...experiments didn’t provide any information on other important factors such as what is the best promoter to use...virus to adjacent areas impacted the experiments we wanted to perform. It required a great deal of time and...Addgene Data Hub, 76% of respondents indicated they wanted to see more negative data (Figure 2.) At the same...help each other by sharing their data. So take advantage of it and get your data - negative AND positive...T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under conditions commonly...Lutomski, C., Jarrold, M. F., Boulis, N. M., & Donsante, A. (2019). Lot-to-Lot Variation in Adeno-Associated...

  10. Getting to Know Expansion Microscopy

    Type
    Blog Post
    Published
    June 20, 2023, 1:15 p.m.
    ...on the Addgene blog Antibodies 101: Introduction to Immunofluorescence  Antibodies 101: Multiplex Immunofluorescence... al. 2022). So what’s a researcher to do if they want to understand the ultrastructural relationships ...ExM protocols are compatible with conventional antibodies, chemical dyes, and genetically encoded fluorescent...integrity of different targets and labels. You may want to check out a few different protocols to look for...et al (2015) Resolving bundled microtubules using anti-tubulin nanobodies. Nat Commun 6:7933. https://doi.org...microscopy using fluorescent Fab fragment secondary antibodies. Methodsx 9:101796. https://doi.org/10.1016/j.mex...labeled using standard fluorescent proteins and antibodies. Nat Biotechnol 34:987–992. https://doi.org/10.1038...

  11. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...Regardless of the type of DNA you want to purify or your motivation of wanting to go kit-less, there’s a method...DNA without a kit. DNA purification kits have advantages: they are convenient and provide uniform, consistent...Cellulose nucleic acid purification Starting material: plant, microbe, blood, or mammalian cells or tissue Product...least entraps nucleic acids. The Botella lab took advantage of this property and developed a protocol that...or even paper towels to purify DNA and RNA from plants, microbes, blood or mammalian cells in 30 seconds...into this mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids...placing the dipstick into a wash solution removes contaminants while nucleic acids stick around. Finally, purified...

  12. Plasmids 101: Colony PCR

    Type
    Blog Post
    Published
    May 12, 2016, 2:30 p.m.
    ...reaction (primers, dNTPs, polymerase) using the supernatant of lysed bacteria as template; and 3) run your...colony PCR primers The first and perhaps most important step to colony PCR is designing primers. There...Blunt end cloning is an example of when you might want to know the orientation of the insert. One member... Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized in the table...clones for later culture: At this point, you will want to hang onto your clones for later use. There are...to start cultures of your positive clones. If you want to store your clones longer term, just streak the... you choose, make sure to use the appropriate antibiotic for selection.   Lysing bacteria and setting...

  13. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ... the presence of oxygen during maturation. This meant that the protein could be taken directly from A....however, point mutations are acceptable. GFP's main advantage over conventional fluorescent dyes of the time...to GFP and new iterations of fluorophores are constantly being engineered. Table 1 lists a few common ...wavelength (blue derivatives) Y145F Increases quantum yield for BFP Y145A and H148D Stabilizes the ...uses currently exist, and new GFP technology is constantly being developed! Fusion tagging: One of the ... plasmid. This can serve as an alternative to antibiotic selection. Plasmids of this type may have the...modified ES cells, which can then be used for implantation and generation of transgenic mice. Purification...

  14. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...are easily studied in living cells using antibodies. Antibodies could further be used to interfere with...screens have been used in several settings, and are fantastic tools for unbiased discovery. With these screens...number of cells needed is lower, and perhaps most important, targeting all genes doesn’t necessarily add value...cytokine known to cause the up-regulation of an important surface protein (e.g. PD-L1 on a cancer cell)....are migratory, and their exact localization is important for their function. Their migration is induced...with the identified surface proteins, and such antibodies could thus be interesting drugs. Known drug targets...in Green Listed.  Provide the list of genes you want to target with your screen in the “Input” box. ...

  15. Typing CRISPR Systems

    Type
    Blog Post
    Published
    March 18, 2025, 1:15 p.m.
    ... simple hierarchy: class to type to subtype to variant. Despite the seemingly complicated steps to arrive...on main function and composition; subtypes and variants are typically based on species of origin. There...currently two classes, six types, 33 subtypes, and 17 variants. Figure 1: Overview of the CRISPR classification...hierarchy flows from Class to type to subtype to variant (shown in italics). Created with BioRender.com.... to as Cascade (CRISPR-associated complex for antiviral defense) complexes, while Class 2 use single protein...after binding to a target. Class 1 is the most abundant of the two, comprising about 90% of identified...others have become more notable in recent years. A variant of V-U, paradoxically called V-K, is a CRISPR-associated...

  16. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...that you want to use for your experiments and verify that your construct is functional. Advantages of the...more about the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway... clone and destination vector carry different antibiotic resistance markers (indicated here by plasmid...coding sequence. If you choose this strategy, it’s important to include the proper protein expression elements...Clone When making the expression clone, it is important to choose the destination vector that best fits...Gateway cloning kit, and MultiSite Gateway plasmids. Constant reading frame When you move a DNA fragment from...stays in frame. High efficiency The positive (antibiotic) and negative (CcdB) selection markers used for...

  17. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    Published
    May 21, 2024, 1:30 p.m.
    ... are more likely to consider cover letters an important part of the application. Even within an organization...core skill, considers cover letters to be very important. On the other hand, our app/dev team rarely, if...that organization or group. This is particularly important for labs and nonprofits, who tend to prioritize...under thirty seconds.” That’s exactly what you would want to talk about! You could describe your organizational... research], and have excellent [list the most important skills for the position]. I am enthusiastic about...about [something that you love and is important for the job]. Notice that the intros above allow you ...include a brief summary of why you are a good applicant. If you haven't talked about it yet, it's also...

  18. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...inconclusive and unreliable. Proper controls provide the constant variables that enable the correct interpretation...of the independent variable you are testing. Importantly, they demonstrate the functionality of your experimental... the transfection reagents and process. It is important to note that optimal experimental conditions, ...Control (mentioned above) could also serve as an important negative control. In this context, the empty vector...interpretation of the results because it provides an important reference point when analyzing the specificity... same time. Although the replicate control is important, it speaks to the consistency of the reagents ...controls are more time consuming and ultimately more important. Ideally, each biological replicate should use...

  19. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...highest quantum yield donor, the highest absorbing acceptor and fluorophores with significant overlap ... theory, is an easy technique, but it is very important to follow some simple rules to avoid common pitfalls...purpose of your studies. These parameters are less important when designing a biosensor, as the distance between...and their spatial orientation will be fixed. The quantum yield of the donor (number of emitted photons per...extinction coefficient of the acceptor (linking the quantity of absorbed light, at a given wavelength, to the...Clover Green Mammalian Expresses Clover (a GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange... mRuby2 Red Mammalian Expresses mRuby2 (a RFP variant) commonly used with Clover pGWF1 Cyan & Yellow...

  20. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    Published
    Aug. 13, 2014, 3:49 p.m.
    ... in March, he and his colleagues took another important step forward by dissecting that S. pyogenes protein...that SpyLigase can be used to link affibodies or antibodies against common tumor markers to subsequently ...cancerous cells expressing low levels of tumor antigen. I asked Howarth to tell us more about SpyLigase... to another peptide tag. Addgene: What is the advantage of peptide tags for building larger protein assemblies... shouldn’t get in the way much. Particularly we wanted to be able to build assemblies through covalent...stability really mattered for efficient capture. We wanted to be able to build a highly multivalent assembly...approaches to improve CTC capture, but I hope that our antibody tentacles may have an impact on this challenge...
Showing: 301 - 320 of 889 results