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We narrowed to 939 results for: OMP

Showing: 521 - 540 of 939 results

  1. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ... an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused to an RNA adenosine...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

  2. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    ...this situation: You’ve spent hours scouring a company’s LinkedIn pages, given yourself the bathroom mirror...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...the candidate has what it takes to thrive at the company and add value, not just “do the job”. When they...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...

  3. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...system, the possibility of creating a replication-competent virus via recombination between the transfer and...recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition...Lentiviral barcoding libraries have been used for complex applications like lineage tracing in heterogeneous...

  4. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money... few ways you can do this. If you are going to complete your colony PCR analysis in the same day, you ... product size on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel... larger sized products for positive clones (+) compared to negative clones. Finally, orientation-specific...

  5. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...is ideal for tracking large collections. Many companies, such as Thermo Fisher and Micronic, provide 2D... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...specific barcodes etched or printed onto them by companies such as Tubewriter. Any lab can benefit from using...tubes themselves. Although we’ve developed our own computer system for tracking samples, it can be done easily...

  6. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...survive as a researcher because of the tough competition for academic positions and poor grant situation...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...

  7. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ... Figure 2: MoClo assemblies depend on compatibility of four-base overhangs flanking each genetic...To combat this, you can increase the volume of competent cells you plate after each transformation and/...high-throughput, we recommend transforming into chemically competent cells in a 96-well PCR plate. Always confirm your... MoClo user, but beginners or those performing complex assemblies should screen a minimum of 8–10 colonies...

  8. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ... and the complexity of effectively deploying them in the lab somewhat mirrors the complexity of understanding.... Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both to..., and jGCaMP8 viral preps distributed per year compared to other AAVs preps from 2016–2023. B) Inset showing...

  9. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...virus as self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...virus’s most abundant proteins and a critical component for viral genome packaging. Addgene recommends...

  10. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Host range depends on whether the host has the components to recognize the ori and allow for replication...organisms. pBBR1-based plasmids and plasmids in incompatibility groups N, P, Q, and W are known to have replicons...list of the bacterial species these plasmids are compatible with.  Table 1: Non-exhaustive list of common... and Srivastava 2013. Family Replicon Compatible with Shown to work in IncN pCU1 Gram-negative...

  11. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...

  12. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn...variants that reached the CNS following IV injection. Compared to AAV9, PHP.B has a >40-fold increase in CNS ...rat CNS: AAV PHP. EB vector dose–response and comparison to AAV PHP. B." Gene therapy 25.5 (2018): 392...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

  13. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...are needed to deliver all the protein and RNA components, and several sgRNAs must be screened to identify...eBook Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

  14. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...target provided that (1) the sequence is unique compared to the rest of the genome and (2) the sequence...for the canonical S. pyogenes PAM sequence NGG compared to wild-type SpCas9, which also displays some ...CjCas9) is even smaller than SaCas9 and is also compatible with AAV delivery. NmCas9, another small ortholog...(2018) Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 ....

  15. Hot Plasmids - August 2020

    Type
    Blog Post
    ...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate...

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

    Type
    Blog Post
    ...meets two conditions:  1) The sequence is unique compared to the rest of the genome.  2) The target is present...found. If you’re lucky enough to find a SpCas9 compatible PAM at your site of interest, that is the most...expanding number of Cas9s with looser PAM requirements compared to SpCas9. xCas9 has a PAM requirement of NG, ... proteins. The Synergistic Activation Mediator complex, generally the strongest activator, consists of...

  17. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...

  18. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136...Kadoch C (2018) Interrogation of Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale...

  19. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...the production of lentiviral vectors that are competent for one round of infection (AKA transduction in...progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with...double-stranded DNA transgene is incorporated into a large complex of both viral and cellular proteins and trafficked...titering IDLVs include detecting vector particle components or performing quantitative PCR titration of vector...

  20. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...genetically encoded tag and exogenously added components. For a review see Lotze et al. (2016). To date...is used to excite GFP, may also excite several components that are naturally present in cells or media....
Showing: 521 - 540 of 939 results