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We narrowed to 629 results for: ncl

Showing: 101 - 120 of 629 results

  1. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...made smaller. Previous attempts to “shrink” Cas9 include the use of St1Cas9 (~3.3 kb) from Streptococcus... approximately 1350 amino acids in size, which includes SpCas9, and shortern orthologs approximately 1000...in mouse liver.  Two other smaller Cas proteins include Cas12b and CasX. While there are no papers yet ...capabilities of full-length SpCas9. This set of plasmids includes AAV-Cas9C-VPR for targeted gene activation. Split...

  2. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...domain - VP64.  Later generations of activators include the SunTag system which facilitates recruitment...Addgene’s website. Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB fusion, both targeted...targeted to the promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors...expression in different species and cell types including (but not limited to) bacteria, yeast, plants, ...

  3. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...but some examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place...resolve these issues is with electrophysiology. Concluding thoughts and moving beyond the mouse While it...applicable to the rat [6]. Technical differences include the ferrule implants and patch cables. One huge... at the Rockefeller University.  His interests include the role of learning and memory in opioid addiction...

  4. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...Several factors contribute to the popularity of GFP including (i) fast and complete maturation to functional...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley et al... conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa = 3.1) and... or media. These sources of ‘autofluorescence’ include riboflavin in the medium or flavin containing proteins...

  5. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...depend on target and sample type. Example steps include: permeabilization, antigen retrieval, and sectioning...available, and your experimental goals. Example steps include: counterstaining, mounting sample on slides, and...you optimize your protocol, always remember to include controls to show that your antibodies are binding...antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share some of their...

  6. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 2:15 p.m.
    ...AAVs to broadly spread throughout an organism, including difficult-to-target tissues like the central nervous...and so on, although we often use abbreviated nomenclature (AAV1, AAV2, etc.) to distinguish the capsids...AAV pseudotyping. It has broad tissue tropism, including liver, muscle, lung, CNS, kidney, retinal, and...serotypes used to target the central nervous system, including neurons, AAV5 is generally the best choice for...

  7. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...for CRISPR genotyping. The most common options include mismatch cleavage assays, such as Surveyor™, T7E1...information profile regarding putative off-target sites including both the edit rate and the repair product sequence...algorithms do not take into account other elements, including those relating to DNA structure (e.g. epigenetic...options exist for unbiased off-target detection, including Digenome-seq (Kim et al. 2015) for in vitro analysis...

  8. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...afflicted by pervasive off-target effects. This conclusion was based upon numerous phenotype discrepancies...importance of using orthogonal methods to confidently conclude the function of a gene of interest. Tools for ...discussed below). Examples of these constructs include pGAPi, which allows easy creation of long hpRNA...experiment are now available on Addgene. These include the construct that allows for easy insertion of...

  9. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...various sources that are difficult to trace. These include the laboratory personnel, the serum, the cell culture...cross-contamination and due to poor lab techniques. These include reusing pipettes for multiple cell lines rather...Contamination! Tissue culture media often have indicators included in the media that turns yellow from lowered pH... periods and affect the total cell viability. Conclusion To sum up, it is always important to prevent ...

  10. Finding Your Science Policy Path

    Type
    Blog Post
    Published
    Sept. 22, 2020, 1:15 p.m.
    ...organizations in a variety of science policy roles including fellowships, volunteer positions, and part-time... with a large set of other societal pressures including economic, cultural, and political. Science policy...science blogs. Other avenues for skill development include leadership of extracurricular organizations, working...opportunities. NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab...

  11. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ... molecule at all, leading to inconclusive results, erroneous conclusions, and wasted time. In fact, many...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from...specifically to the expected protein. Common approaches include using RNAi to knock-down expression of your target...

  12. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ..., introduced the “Three Es of Immunoediting” nomenclature. 1. Eliminate - Latent tumors are recognized...on immune system Conventional cancer therapies include radiation therapy (owing to contributions from ...blocking. Other molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific... factors researchers are beginning to focus on include: Cell heterogeneity Tumors are composed of a wide...

  13. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ... Figure 1: Labeled diagram of an antibody including Fc, Fab, heavy chain, light chain, constant region...antibodies for experiments in a variety of fields including neuroscience and immunology. They can be used ...analyzed using techniques such as flow cytometry. Conclusion Whew! Now you know that antibodies can be used...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ...

  14. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...for plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!   ...destination vectors. Destination vector options include humanized wt Cas9, dCas9 (transcriptional repressor... Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable ...filler sequences. You also have the option of including a previously-designed tyr gRNA, which causes hypopigmentation...

  15. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    Published
    May 27, 2025, 1:15 p.m.
    ...sneak in from almost any step of the process and include things like genomic DNA, RNA, proteins, salts, ...sustainable practices for the lab. Plastic consumables (including pipette tips, plates, and yes those purification... kits from a variety of manufacturers. It also includes a rough comparison of list price between these...Yeast, BAC, transfection-grade While we’ve included a sample of some commonly used kits, please note...

  16. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    Published
    Feb. 13, 2014, 1 p.m.
    ...for plasmid. pXXXXX-XXXX Step 1: Backbone name Include the empty backbone name in your plasmid name. This...backbones, visit Vector Database. Step 2: Insert name  Include information about the insert in your plasmid name...contains a mutation or modification, this should be included in the plasmid name. Mutations are generally listed...

  17. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...the template. The PE3 system does just this by including an additional sgRNA. Using this sgRNA, the prime... number of unintended prime editing outcomes, including indels. Therefore, they engineered a dominant-...     ✓ ✓ reflects an innovation always included in the indicated tool. ~ reflects an innovation... strategies in our CRISPR guide. Some examples include: TwinPE: two prime editors make complementary ...

  18. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ...species, antibody species, or whether the results include knockout data.    Figure 2: Search options in ...description of the experiment. If that data was included in a publication, you'll find that information... is easy!  Here at Addgene, we love all data, including negative data, which can help researchers save...

  19. What's New in CRISPR - May 2019

    Type
    Blog Post
    Published
    May 28, 2019, 12:49 p.m.
    ...per gene and includes ~10,000 gRNAs targeting intergenic regions. The libraries also include 2,400 gRNAs...recently deposited to Addgene and you’d like it to be included in the next What’s New in CRISPR blog post, please...

  20. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    Published
    Feb. 25, 2021, 2:15 p.m.
    ... to a post-academic career. This training was “included” with their tuition, so to speak.  Unfortunately...the development of priorities for pd|hub which include building capacity to identify the most effective...scientists, but it is essential that we integrate inclusive career readiness training in this educational ...
Showing: 101 - 120 of 629 results