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We narrowed to 798 results for: REP

Showing: 241 - 260 of 798 results
  1. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...chromosome. During DNA repair, there is a chance that the two breakpoints will repair without the section... a derivative of Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors...template strand left behind that the cell can use to repair. Figure 3: General overview of the Cascade-Cas3...ssDNA past the nuclease domain, Cas3 is able to repeatedly cut the DNA at an efficient rate. These cuts ... or just random mutations if the chromosome is repaired with all the original DNA. With Cas3, you don’...References Loeff, L., Brouns, S. J., & Joo, C. (2018). Repetitive DNA reeling by the Cascade-Cas3 complex in nucleotide...
  2. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ...the new plasmids, antibodies, viral preps, and more in the repository through our Hot Plasmids articles....vivo expansion cultures from days 7, 14, and 27. Reproduced from Singh et al. (2025) under CC BY license.... available with T-Sapphire, EGFP, or mScarlet reporters, so you can choose a color compatible with the...and AAV11 tropism and applications. A) AAV6 is reported to have broad tissue tropism, including skeletal...hematopoietic cells, and immune cells. B) AAV11 is reported to target skeletal muscle, kidney, spleen, lung...the Cre recombinase payload activates a floxed reporter gene (genetically encoded or delivered separately...
  3. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...vector preparations, we have used quantitative PCR to evaluate the physical titer of our preps. We often...often repeat this assay to confirm that our results are accurate. But what if we could repeat this assay... for ddPCR set-up. Prepare the master mix in a separate area from where you prepare your sample dilutions...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...
  4. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ... an AAV vector to express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes...dopamine beta-hydroxylase (DBH, magenta) to label norepinephrine (NE) expressing neurons and GFP (green) to ... right AAV for your system or to successfully replicate published works. Of particular importance to these...experimental settings. It is a curated set of technical reports on AAV vectors from Addgene and the CLOVER Center...Center, each one submitted by researchers like you. Reports are reviewed by Addgene scientists for clarity ...performance. Yet, most of the technical data needed to replicate and build on others’ results is available not ... in Adeno-Associated Virus Serotype 9 (AAV9) Preparations. Human Gene Therapy Methods, 30(6), 214–225....
  5. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ..., Zetsche et al. hypothesized that Cpf1 might represent a functional CRISPR nuclease that could be adapted...about the same size as Cas9’s, but with the direct repeat preceding the spacer rather than following it. ... only a short stem-loop structure in the direct repeat region is necessary for cleavage of a target. Cpf1...are difficult to modify through homology-directed repair (HDR). Cpf1 also expands the number of sites that...experimentally confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions...
  6. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...these proteins, as well as the viral long terminal repeats (LTRs) are different. As a result, lentiviral and...adenoviral protein E1A, which has been shown to repress expression of HIV-1 LTRs (more information is available... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...all) Addgene transfer plasmids are replication-deficient meaning they can be used to create virus that is...Which genes are deleted or modified to result in replication deficiency? What percent of the vector components... vectors are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential...
  7. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...express my concerns and request a replacement. I got my replacement in the end and the issue was resolved... A degree in the life sciences prepares one for numerous non-academic careers. Still, many start their...support roles. Examples are Technical Service Representative, Technical Support Specialist, Technical Support...less suitable for troubleshooting or issuing replacement products. In order to find good and helpful answers...certainly there won’t be a shortage of amusing reports where someone tries to “optimize” a particular ...job. Solutions may also be achieved by simply replacing the product in question. This can solve the problem...
  8. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ... organisms) -  perhaps you’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project...used to study aspects of cognition. Spiralians - Crepidula atrasolea (Jonathan Henry) This black footed slipper...real bottleneck. New gif for #MolluscMonday: #Crepidula atrasolea juvenile, light microscope with polarizing...it in a fertilized egg to get edited embryos (replacing the nucleus is still very difficult). Cuscuta ...targeted gene mutations in anoles lizards or in any reptiles at all. But new success is coming with CRISPR/...full hydra and hydra have asexual and sexual reproductive life cycles. One can inject plasmid DNA in the...adult female mosquitoes. Now they are trying to replace embryo injection (hard to do, expensive, not every...
  9. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...different CGBE tools allow the researcher to direct the repair of the abasic site. But what if an editor could...excision is the most common type of base excision repair. But mammals also have an enzyme to remove inosine...collaborative team from the David Liu and Dali Li labs reported a similar tool shortly thereafter, using mouse... or to create the substrate for a base excision repair pathway. The team behind AYBEv3 introduced the ... the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template —...editors Excises G, T, or C, respectively, to be replaced with another base. References Chen, L., Hong...Transversion Editors Read about four base editor reporters Resources on Addgene.org Browse CRISPR plasmids...
  10. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...jejuni (Cj) NNNNRYAC Streptococcus thermophilus (St) NNAGAAW Treponema denticola (Td) NAAAAC...in mammalian cells. The REPAIR (RNA Editing for Programmable A to I Replacement) system fuses Cas13b to...when trying to edit a gene using homology directed repair, since HDR-mediated gene editing is most efficient...Hu et al., 2018).  In 2020, the Kleinstiver lab reported the development of near-PAMless Cas9 variants....below.  Cas9 species PAM sequence ’) Streptococcus pyogenes (Sp) 3' NGG Staphylococcus aureus...
  11. New Viral Vectors - Spring 2025

    Type
    Blog Post
    Published
    April 29, 2025, 1:30 p.m.
    ... to conduct experiments faster, skipping the preparation steps of production, purification, and titering...customizability, choosing the cargo, serotype, and prep size they need for their experiments — and we’ve...them out in record time.  However, our in-stock preps remain a popular choice thanks to the convenience...cortical interneurons. Take a look to see if these new preps might enhance your research!   Table 1: New ready-to-use...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV-CAG-emiRFP670...help, with over 100 user-submitted viral vector reports. And don't forget to use the link below to keep...
  12. New Viral Vectors - Summer 2024

    Type
    Blog Post
    Published
    July 9, 2024, 1:15 p.m.
    ...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 20... 20 additions we've made to our viral vectors repository since March.  Addgene is now offering an AAV2...gene expression! We've also added AAV1 and AAV5 preparations of several variants of a new optogenetic tool...Yizhar lab. Addgene is now offering AAV1 and AAV5 preparations of several variants of a new optogenetic tool...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link...
  13. Deaminet 2026: Breakthroughs in Base Editing, Deaminase Biology, and Therapeutic Translation

    Type
    Blog Post
    Published
    March 10, 2026, 1:14 p.m.
    ...Mechanism of DNA replication fork breakage and PARP1 hyperactivation during replication catastrophe. Science...of CRISPR tools, with base and prime editors representing two of the fastest growing segments of the collection...domain in dark red and the docking domain in pink. Reproduced from Arantes et al. (2024) under a CC BY-NC 4.0... Figure 3: Labeling of Cas9-mediated R loops. Reproduced from Wang et al. (2025) under a CC BY 4.0 License...strategy. Figure 4: Schematic of PERT. Reproduced from Pierce et al. (2025) under a CC BY 4.0 License...(UC Irvine) showed that APOBEC3B activity at replication forks induces APE1-dependent abasic site cleavage...therapeutic strategies. Sarat Chandarlapaty (MSKCC) reported that APOBEC3-dominant breast tumors resist endocrine...
  14. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ...gradients? Excellent! Now, let’s head west, intrepid reader! Protein Prep The first step in any western blot is...minutes) at 37 °C, if you suspect aggregation in your prep. If your proteins are the wrong size on your gel...will simply load an extra ladder or technical replicate (ie, leftover sample) to fill the relevant wells...antibody’s dynamic range. A) The protein dilutions, representing expected concentrations, are all within the ...antibody’s dynamic range. B) The protein dilutions, representing expected concentrations, are outside of the ...blot, you may want to consider using a different reporter or, if you don’t need to compare the protein amounts... amounts directly, stripping and reprobing. Just make sure to go from least to most abundant protein if...
  15. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...fluorophore-conjugated streptavidin protein, which binds the primary antibody through the biotin-streptavidin interaction...detection. The advantage of this system is that streptavidin conjugates are commercially available for a ...fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids at Addgene!   ... two-protein labeling by utilizing(a) biotin-streptavidin binding and (b) antibody host species interactions...include transcription factors, histones, and DNA repair enzymes. Note that when using the nuclear fix/perm...or protein-protein interactions, like biotin-streptavidin binding and antibody host species reactivity...
  16. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    Published
    Aug. 23, 2016, 2:30 p.m.
    ...talk to the occasion Don’t just plan to reuse a prepared talk unless it perfectly fits the occasion.  I...figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until ...for poor quality slides with sufficient time to prepare (and it does take time). For important talks, ask...loves a great animation. 8. Predict questions, prepare answers Speaking pros always have a few extra slides...come up with clear and concise answers. This preparation will help you avoid giving only yes or no answers...their phones. Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than...when I then ask “and who has deposited to the repository?” When most or all of the hands go down it is...
  17. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    Published
    June 26, 2017, 1:31 p.m.
    ... are overrepresented on Twitter Mathematical, life, and physical scientists are underrepresented. It’s...interested in getting started? Reply in the comments below or tweet us your reply & #scientistswhotweet. And...users. Biologists rank #8. Figure 1: Highly Represented Scientific Disciplines on Twitter. Twitter users...Follower Network of Scientists on Twitter. Each dot represents a scientist. Only scientists who follow each ...
  18. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...engineered not to cleave DNA and instead activates/represses gene expression by various mechanisms. The RNA...cleaving of DNA occurs, not for deadCas activator/repressor experiments where only localization of the Cas...will generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation...insertion of DNA by the NHEJ pathway during DNA repair of the break to allow for site-specific detection... these approaches is that NHEJ must perform the repair to allow for detection, not Homologous Recombination...non-cutters If your Cas experiment involves activation/repression or any other effect which does not depend on ... be an appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would also identify...
  19. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    Published
    Sept. 12, 2023, 1:15 p.m.
    ...data repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for ... Calculator app for iPhone Reagent and Data Repositories These links can help you find experimental and...for Cas enzymes.  Open repository of CRISPR screens Cellosaurus: A knowledge resource for cell lines IBEX...IBEX Knowledge Base: A data repository for validation data on IBEX reagents.  mAb3D atlas validated antibodies...human, and marmoset brain IHC.  FPbase: A data repository for fluorescent proteins and their properties...
  20. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ...the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think...pitch a Hot Plasmids section (antibodies and viral preps welcome!) here. Here's what you'll find in this...Liu Z, et al. Cell. 2022.  Find JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu...adapted from Govorunova et al. 2022.     HcKCRs represent a long-sought tool for researchers to control ...s41593-022-01094-6. (This work was also shared as a preprint: https://doi.org/10.1101/2021.09.17.460684.)   ...
Showing: 241 - 260 of 798 results