We narrowed to 897 results for: ANT

...CRISPR activation uses dCas9, a CRISPR protein variant lacking its endonuclease ability, to bind to genes...themselves. SunTag, SAM, and VPR have all shown significant improvements upon the initial dCas9-VP64 method... this method is its construction: it relies on antibody chains, which are relatively large and are not...Performance Generally, VPR is found to have significantly higher activation levels than the initial dCas9...activation levels are similar to that of SunTag. An advantage to this method compared to other notable CRISPR...

... would be infected by a lab-grown virus, it's important to understand the risks of the specific virus ...necessary step during production), there is a significant risk for aerosol contamination. As most centrifugation...ethanol (other surfaces). For AAVs, which are resistant to ethanol and isopropyl alcohol, 0.5% peracetic...sharps procedures for your organization will significantly reduce the risk of an accident (the most common...more likely to make a mistake and more likely to want to cut a few corners. For many researchers, viruses...

...Outbred strains of mice are useful for comparing dominant and recessive traits and their impact on diseases... case, a positive selection marker such as an antibiotic resistance gene is also included to select for... are injected into a blastocyst which is then implanted into a female animal for gestation. Nucleases ...are microinjected into a zygote, which is then implanted into a female mouse for gestation. All three can...nucleases to generate an engineered mouse model has advantages over the injection of a targeting vector alone...

...advocacy skills “bootcamp” for a focused group of participants. More information can be found at http://futureofresearch.org...graduate students who work as teaching or research assistants to join unions. The topic of unionization continues...productive routes for advocacy efforts and help participants develop the efficacy of their advocacy skills...of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University of Massachusetts... Developing effective science advocates is an important way to push the balance from challenge to opportunity...

...strength as a determinant of gene expression levels, the terminator also plays an important role in RNA ...expression vectors use double terminators to reduce unwanted translation of downstream elements. A high affinity...polyadenylation. The addition of the poly(A) tail is important for stability of the mRNA, protection from degradation...Maitland NJ. An Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered...

...at my workshops, I get nods from nearly every participant. Robert Bramson, author “Coping with Difficult...people: Hostile-aggressives, who are usually antagonistic and impolite, Complainers, who spend most ...people. Some students and postdoctoral fellows are hesitant about expressing their ideas, especially if their...solutions that will meet both of your needs. 6) Put important agreements in writing One of the most frequent...from your supervisor. Whenever possible, discuss important issues in person or over the telephone instead...

... in high-throughput libraries My labmates and I wanted to find a way to predict the efficiencies of pegRNA...Figure 1). Through this method, we found key determinants of prime editing efficiency such as the length...et al., 2024) have highlighted how chromatin substantially affects prime editing outcomes across many genomic..., A., Kuzmin, I., Raudvere, U., Peterson, H., Liberante, F. G., & Parts, L. (2023). Prediction of prime...efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org...

...regulation, but enhancers and chromatin looping play important supporting roles. Enhancers are cis regulatory...ligation creates chimeric circles of DNA that are distant linearly but close together in space. The chromatin...interactions. The chromatin story told by CAPTURE 2.0 is important, but it’s not the only plot line in gene regulation...CRISPR users Learn how to Choose the Best Cas9 Variant for Your Next Experiment Learn More About Designing...Additional Resources on Addgene.org Find Other Cas9 Variants for Purifying Genomic Regions of Interest Here...

...traveling for campus visits. This is definitely an advantage of surviving in a virtual world over the course...Having notes close to your computer with things you want to highlight in the interview is another way to ...staff have become a new way to allow you to get important information about the campus even if you can’t...and the in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity...exception. However, using the virtual world to your advantage can strengthen your application and help you decide...

...design, batch primer design or batch analysis. We wanted to relieve scientists of the burden of having to... to analyze  CRISPR repair outcomes, which is important for quality control. The only ones we found that...that worked with NGS data were: Crispresso CrispRVariants AmpliCan We chose to adapt Crispresso by Luca...their "summarisation score".) For CrispyCrunch, we wanted to consider both factors, as a bench scientist ...coding region in a UTR, CrispyCrunch flips the two important base pairs of the PAM (NGG -> NCC, CCN -> GGN)...

...gRNAS1, and pAdSh.U6.gRNAGFP. Gonçalves says that advantages of AdVs include their episomal nature and very...efficient introduction of DNA into therapeutically relevant, non-transformed mammalian cells. These viral ...Disease, Duchenne Muscular Dystrophy, Alpha1-antitrypsin deficiency (most widely used so far) or for cancer...used, the CRISPR approach based on HDR has the advantage of being a potentially universal treatment (Stephens... could be realized via simple switching of the relevant gRNAs and donor DNA Adenoviral CRISPR/Cas9 genome...

...below). Let’s take a look at the features and advantages of the optimized human CRISPR libraries. Brunello... enough for efficient library performance, an important criterion for probing a limited number of cells..., confirming again that sgRNA design plays a significant role in library efficiency. Try the Dolcetto ...al. demonstrated that the Calabrese library substantially identified more hits than the SAM library approach...Libraries available at Addgene Learn about the advantages of pooled CRISPR libraries for large-scale functional...

...and reported results are encouraging. Importantly, a significant amount of basic research is being done...that can be monitored non-invasively over time. Importantly, it is an immune privileged compartment, meaning...meaning that it can tolerate the presence of antigens (such as viral vectors) without triggering a destructive...improve the technology: engineering of new capsid variants, designing expression cassettes to regulate and...

... your research. RNA editing advantages RNA editing has multiple advantages over more traditional DNA editing...stretches of duplexed RNA. Using an ADAR2DD catalytic mutant, they showed that editing is mediated by ADAR2DD...precise than shRNA knockdown, it still displays substantial off-target activity. By mutating residues in ...discussed above, the ability to edit RNA has multiple advantages, including reversibility and use in non-dividing...

...months, pausing or terminating employment for a significant fraction of the workforce. Wet lab scientists... open-ended question is difficult to answer quantitatively without broad, unbiased data. Therefore, we...cases being reported (right, red), likely due to quarantining {% video_player "embed_player" overrideable...exponential decay model, with dips during weekends and significant holidays. Figure 4: Plasmid request trajectories...parameters from relatively sparse data from China. An important additional caveat is that our dataset contains...

...learning experiences happened in the lab, and he wanted to provide the same kinds of opportunities for ...interaction with other proteins.   It is important to note that research activities in a CURE must...hypotheses being rejected, etc.), while reminding participants of the value of this approach and encouraging...hMDH1). They then expressed the original and recombinant enzymes in bacteria, purified them to near homogeneity...by making a greater number of students active participants in the research process – so if this sounds ...

... should not even be the real goal. We are all constantly negotiating our energy and attention. The goal...you’ll need to care for an elderly family member, want to train for a triathlon, finally buy that charming... someone there clapping more often than not. You want your family and friends to know you care, but they...they need to know that you have an important life too. They will respect you if you show respect for your...You can handle that, right? Big tip: this is an important topic to discuss and agree on before you get enmeshed...

...were less significant than when non-targeting guides are used as controls. Less significant p-values lead...genome-wide screens.   Figure 1: Summary of the advantages of the new approach to genome-wide CRISPR screen... Screens Before diving into the results, it’s important to note that in Morgens et al., guides were considered...of on-target activity. However, there was no significant difference in depletion of ricin sensitizer genes...

...External collaborations are becoming increasingly important as more complex approaches to scientific questions...collaborations and partnerships are also very important in a non-research context, especially when considering...initiative to find a suitable partner, connect the relevant teams, coordinate implementation, and now must...start and maintain external collaborations is important? In my current job at Addgene I work with people...work with others outside your organization is important for success in research-intensive careers. In ...

...thought to activate programmed cell death or a dormant state for bacterial cells that have been infected... Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They characterized...displays similar knockdown efficiency but no significant off-target effects, a distinct contrast to the... In short, the Cas13a molecule brings a much anticipated element of diversity to the CRISPR toolbox and...Check Out Our CRISPR Featured Topic Page Learn about Anti-CRISPRs Multiplex Genome Editing with Cpf1 Resources...

...viral genome and the absence of unexpected DNA contaminants. The data is also used to determine recombination...properties of the final vector preparation, the quantity of gram-negative bacterial endotoxin is ensured...within representative vector preparations was quantified with electron microscopy after negative staining...Approximately two weeks post-thaw, cell culture supernatant is tested for mycoplasma contamination. To date... Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors 101: An Introduction to AAV References...

...detectable non-specific activity. Its major advantage is the significantly reduced amount of off-target toxicity...enzyme can lead to off targets and result in significant, negative effects in RNA targeting experiments...using Prime editing alongside some creative (and significant!) protein engineering. They combined Prime editing...The potential of CRISPR has yet to fully taken advantage of, but exciting new tools like these represent...

... comments on the AMA process below, but we also wanted to share some thoughts on why we decided to do ...Software for Biology) on Reddit and got some substantial traffic that we thought it would be a good idea...worried about selling yourself and really do just want to get information about yourself and your work ...internet content for scientists in training is abundant, helpful, and sometimes excellent. Finally, as...as I said in the AMA, it’s incredibly important to find good mentors and incentivize good mentorship: ...

...volume of lower titer virus. In such cases you may want to consider concentrating your viral prep. Storage...al. found that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, ...protein attaches to cell surface receptors, other important binding events take place during the course of...source. Labs that routinely produce lentivirus may want to consider testing a variety of cation and serum...Ward T.H., Margison G.P. “Mycoplasma infection significantly alters microarray gene expression profiles.”...

... out of fashion but is still useful and most importantly, effective. In the research world, there is a...plasmids The design of shRNA plasmids requires three important steps: Designing the shRNA sequence Choosing ...polymerase II promoters. The design of these plasmids is meant to more closely mimic pre-miRNAs, and the use of...19px;font-family: helvetica;","color":"#FFFFFF","variant":null,"font":"helvetica","style":"font-family: ...resources on the Addgene blog RNA Interference in Plant Biology: New Tools for an Old Favorite CRISPR 101...

...but it’s also important not to load yourself up with too many tasks. If you’re constantly switching between... recommend the book, “Bored and Brilliant” for learning more fantastic reasons to remove yourself from...ll discuss in the planning section below, it’s important to dedicate specific amounts of time to specific...

...thanks to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is a PhD candidate...This post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-...cut will continue to flow and be captured by an antibody further along on the strip. Highlights: Of the... cDNA. Isothermal amplification is done at one constant temperature, eliminating the need for thermocyclers...

...should be supplied (Baloban et al., 2017).  It is important to consider effective brightness of NIR FPs in...molecular brightness (a product of the fluorescence quantum yield and the extinction coefficient) (Shcherbakova...They were engineered to incorporate BV. Their advantages are their small size (17 kDa for miRFPnanos, ... highlighted in blue. aExtinction coefficient. bQuantum yield. cDetermined in mammalian cells. dDetermined... near-infrared proteins provide a scaffold for antigen-dependent fluorescent nanobodies. Nat Methods submitted...

...good article at NatureJobs: Interviews: The All-Important Job Talk. 1. Watch other people give great talks...Think Keystone Keynote presentation or Ted Talk. You want to give the audience something to think about, argue... time to prepare (and it does take time). For important talks, ask your peer mentors to check your slides...Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than too long, and...to practice and over time you can master this important science skill. Additional Resources Joanne'...

...CRISPR tools for use in plants.  Listen to the segment on CRISPR tools for plants. Special thanks to our...enhancing iPSC production, and CRISPR tools for plants. You can find additional hot plasmids in our quarterly...: Interview with Darrell Kotton Find additional plant biology blog posts Resources at Addgene.org Sign...