...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...

...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...

...advocacy non-profits like Phipps Conservatory or the Brilliant Club and share your knowledge at local schools...Student Voice are two examples. Many blogs like Plant Science today, Soapbox Science (hosted by Nature...Science, Media and Communication. In the US, UC Santa Cruz, MIT, UW Madison, and Boston University offer...where you may take a science track. If you don’t want to commit time and money to a full-time graduate...communication. If science journalism is the path you want to follow, check out the distance learning courses...

...these communities, which is in turn crucial if we want to make sure that everyone can be informed and empowered...individual consumers of technologies and as societal participants in the political process. My role at pgEd I ...who are interested in such careers should take advantage of their academic environment and attend seminars...either giving or running these events will be important both in preparing you for future work in these...communicate your work to a non-technical audience is an important skill for every scientist. After all, scientific...

...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...

...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...

...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...

...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)...

...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...

...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 ...

...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...

...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...

... 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...

...Protocols Recombinant Antibody Affinity Purification with Protein A or Protein G Recombinant Antibody Affinity...like... Antibody Plasmid Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies...describes how to affinity purify recombinant antibodies from cell culture supernatant using Protein A or Protein...to affinity purify recombinant antibodies. Workflow Timeline Day 1: Purify antibody Day 2 or later: Buffer...Equilibrate filtered tissue culture supernatant containing the recombinant antibody to room temperature (see Transfection... will ensure depletion of the recombinant antibody from the supernatant. Wash the column with 25 mL of...

...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...