...is still important. It needs to express value, clearly communicate exactly what a site offers to their... for users to get where they desired. It became clear to us that we needed to improve search and navigation...as simplifying the page, and making it all look clean and easy to use.  Easily search for plasmids (and... new search bar is now front and center, with a clear call to action - Search! Along with this we’ve also...element. We also set out to make sure the page looked clean, professional, and modern. Instead of trying to ...next science challenge. If you have any feedback, please contact us and let us know how you’re feeling.  ....  Additional resources on the Addgene blog Learn more about Addgene's new search tool Read more about...

...protein’s reading frame, leading to the production of a smaller, but at least partially functional protein...individual experiences progressive muscle wasting leading to death at around 30 years of age. Despite the...depositor Charles Gersbach used paired zinc finger nucleases to remove exon 51 in DMD patient myoblasts. They...force, and other muscle tests, muscle function is clearly improved, although not to wild-type levels. With...that dystrophin restoration is maintained for at least six months. Future directions and obstacles to clinical...through genomic excision of exon 51 by zinc finger nucleases. Mol Ther. 2015 Mar;23(3):523-32. PubMed PMID:...PMC4393360. Additional Resources on the Addgene Blog Learn How CRISPR Could Be Used to Treat Cataracts and ...

...enzymes. Note that when using the nuclear fix/perm buffer to label intranuclear proteins of interest, you can... ligases; and many more. This process, however, leaves the nucleus intact. If markers of interest are ...can label cytosolic proteins alongside the intranuclear ones. You do not need to use a separate fix/perm...typically used. Inside the cell, the ester group is cleaved off by enzymes, turning the resulting molecule ...You made it through this blog post and hopefully learned a few new methods of target visualization beyond...specific downstream applications, while maintaining a clean cell surface, dump gating is a useful way to get... labeling can be performed on cytosolic and/or nuclear markers. Lastly, stains, based not on antibodies...

..., because leaky expression of toxin could select for mutations in the toxin itself (leading to counter-selection...gene with an antibiotic resistance cassette. This leaves a “scar” in the chromosome, sometimes interfering...ways to create scarless mutations, ones that don’t leave any undesired scars in the chromosome. These methods... The resolution step (“double crossover”) which leads to the desired mutation and excision of the vector... paper. We selected the three toxins so that at least one might be useful in a broad array of pathogenic... switched over to pTOX. However, we feel it has clear advantages, and are hopeful that continued development...Plasmids 101 blog posts Find our CRISPR blog posts Learn more about Lambda Red recombineering Resources ...

...for you, if you can identify them in yourself and learn how to sell them. Types of Conflict Before explaining... on something and a disagreement festers, it can lead to hurt feelings, low morale, poor decision making...situation. If left unchecked, this type of conflict can lead to people disagreeing with each other just for the...the real source of the conflict. It can be easy to leap to the conclusion that a conflict is personal (”... listener, but I aspire to  be. If you don’t at least aspire to be a good listener and consciously practice...wanted. However, a poorly brokered compromise could leave everyone feeling vaguely unsatisfied. In arbitration...than either A or B, but the two parties will have learned that by working together, they can create unique...

...its endonuclease activity to generates a ssDNA substrate which is then nicked by its endonuclease domain...engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about AAV...of Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins share a common...virion assembly, the role of MAAP has not been clearly defined yet. Now you’ve got a basic understanding...AAV RepCap plasmid! Check out this blog post to learn more about the AAV transfer plasmid! References...

...depurination, the hydrolysis of the glycosidic bond leads to the release of a purine (adenine or guanine) from the...DNA are repaired by base excision repair, but can lead to mutations in single stranded DNA undergoing replication... from E. coli is the presence of the protein endonuclease I encoded by the gene endA. Nowadays, the most...gene has been removed/mutated to neutralize the nuclease activity. For strains that still carry this gene...introduced in the purification protocol to remove endonucleases activity. For these reasons choosing the correct...most downstream processing. On the other hand, nuclease free water is a great option because it provides...

...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...mutation inactivates the RuvC domain, so this nickase cleaves only the target strand. Conversely, the H840A mutation... in the HNH domain creates a non-target strand-cleaving nickase. Instead of cutting both strands bluntly...distances (40 - 130 nt). From these results, it’s clear that editing is much higher in the PAM-out configuration... At the AAVS1 locus, the two nearest gRNAs had cleavage sites 12 nt and 13 nt away from the target site...wide margin, exhibiting >20% repair efficiency. Clearly, a nickase strategy can expand the targetable region...post. Additional Resources on the Addgene Blog Learn about Homology Directed Repair 3 Tips to Improve...

...Careers in Science Communication blog series, you’ll learn about Caitlin Runne-Janczy, a Product Development...While her love of educational science writing is clear, Caitlin’s career plans weren’t concrete when she...what I wanted to do, but knowing that I wanted to leave the bench.” Her path to figuring out her career ...writer.” At the end of her postdoc, Caitlin had a clearer idea of her career goals: “My plan was to go into...for part two of Caitlin’s interview where you can learn what it’s like to work for a science education company...series!  Additional Resources on the Addgene Blog Learn more about alternative science careers here Be like...while in grad school Resources on Addgene.org Learn more about careers at Addgene ...

...non-academic career Learn more about alternative science careers here Resources on Addgene.org Learn more about...series was written for selfish reasons: I wanted to learn about careers in science communication. When I started...away-from-the-bench career option. So if you’re interested in learning more about science communication careers, you’...was a science writer, reached out to me. She was leaving a writing position and looking for someone who ...s paid off. Work is never boring. “I just keep learning on the job. It keeps it interesting.” And while...topic, but I haven’t met any.” A few other tips: “Learn HTML, write a lot to make sure you don’t hate it...

...professional Addgene is a thought leader in scientific sharing, leading by example to increase sharing in...Resources on Addgene.org Learn More About Our Mission Meet the Addgene Leadership Team Get the Latest News...arts organization. Many people are surprised to learn that Addgene is officially filed, recognized and...is a nonprofit? The term "nonprofit" is a bit misleading. Nonprofits can generate revenue (as Addgene ... Xuan Additional Resources on the Addgene Blog Learn about Our Collaboration with Fellow Nonprofit Seeding...

...Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts in your experiments. Read this ...Professor at Albert Einstein College of Medicine, to learn how problematic oligomerization can be and how to...that occur during development. Read this post to learn more about recombinase-based state machines and ... Green (both formerly at the Wyss Institute) to learn how these amazing tools may be used to rapidly diagnose...you can use to study basic virus biology while leaving virulent components behind. Minigenomes are stripped...and transcription as well as reporter genes. To learn how you can use minigenomes in your research, check...

...s antibodies are plasmid-based and recombinant. Learn more about recombinant antibodies here. Finally,...Finally, Addgene is a nonprofit that connects colleagues and is invested in improving research infrastructure... to maximize the reach and impact of your work. Please email [email protected] if you would like to ...plasmids via Addgene! Email [email protected] to learn more or get started.  If you have suggestions about...ideas for new materials to add to the collection, please let us know by filling out this suggestion form... will find useful antibodies for your research. Please consider sharing your experiences with antibodies...Resources We would also like to acknowledge our colleagues at the Developmental Studies Hybridoma Bank, ...

...nature of non-homologous end joining (NHEJ) often leads to the generation of indels and thus frameshifts...activation) and CRISPRi (inhibition) technologies – a nuclease-dead Cas9 (dCas9) is directed near the promoter...landscape of mismatches that can nevertheless still lead to activity has been established, and can be used...that the consequences of off-target activity, at least for the experimental conditions used, led to no ...the generation of truly isogenic cell lines than nuclease-mediated off-target effects.” Further, large-scale...al., 2019). Again, the experimental strategy is clear: for any gene of interest, one should require that... Liu DR (2020) Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....

... about roles in this field, most jobs needed at least 3 months experience working in a developing tropical...positive experience. I enjoyed helping people and learning to explain concepts in ways that people from another...traditional medicine couldn’t fight effectively. I had to learn how to cope with a different climate and culture...work became monotonous very quickly; as soon as I learned how to do the different tests and became efficient...where one drug does not work for all patients, leading to very high mortality. I really loved the fact...cannot be as adaptive. I was also surprised to learn that I enjoy the customer service side of my job...vital because otherwise I’d lose touch with my colleagues and have misunderstandings.  Working long distance...

...the Addgene Blog Learn more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent...brightest since it had an emission near 600 nm – a clear demonstration of why a BRET reporter can improve...Many intracellular sensors of events like calcium release have been made using fluorescent resonance energy... molecules. FRET sensors face challenges of photobleaching, autofluorescence, and, in the case of exciting...wavelengths, excitation of the optogenetic regulator leads to spurious activation of the FRET sensor. Researchers...et al. (7) wished to trigger intracellular Ca++ release with an optogenetic receptor, melanopsin, requiring...#83926). Live-Cell Voltage Sensor  Ingaki and colleagues (8) were interested in using photoactivatable...

...bound lipoproteins Article contributed by Leah Schwiesow A leading cause of death worldwide, cardiovascular...ever like to write about a recent plasmid deposit please sign up here.        A retroviral mouse CRISPR...genetically. To solve this issue, Sarah Teichmann and colleagues created a retroviral CRISPR knockout library ...lipoproteins through the circulatory systems, but can also lead to the formation of atherosclerotic plaques. ApoB...through triglyceride/cholesterol does not provide a clear view of either of these metrics (Sniderman et al...-LP size and concentration, Steven Farber and colleagues at the Carnegie Institute for Science have developed...

...of a paper is the one that you need to spend the least time on—just put down a basic outline and refer ...figure them out. But, when they come to light, it’s clear that what the researchers thought was a problem ...role, and we take our responsibility to push for clear communication of science very seriously. That’s ...and transparency of methods sections, including clear information about reagents, and why other publishers...want to be? The part that is still missing is a clear alignment of incentives. Right now, researchers ...evidence of sharing for grant renewal—but there is clearly room to do more. If we could really instill a culture...Her current responsibilities encompass overall leadership of the Cell Press editorial department, a team...

...often a hard question to answer if you plan on leaving academia, but don’t quite know what you do want...goal, and when you realise that your goal might not lead to a career you actually want, you can feel lost...Journal Editor Science Marketer Software Developer Learn Science Communication Skill from a Student Group...in the media and in films, which can often be misleading. Key aspects to determine are working hours (... the job require public speaking? Will you be leading a group or working as part of team? Will you be...application. Additional Resources on the Addgene Blog Learn How to Experiment with New Careers while in Grad...Koeris' Journey from Grad Student to Entrepreneur Learn to Network and Make the Contacts for Your Dream ...

...miscommunication, a lack of leadership, and inappropriate tools or writing approaches can lead to frustration, delay...Weill Cornell Medicine. Discussing data with colleagues can be more difficult during the pandemic era...can contribute and sketch on the board without leaving Zoom. The annotate function can also make the screen-sharing...cultural backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and...ways of working, reconnect as human beings and colleagues, eventually encouraging purposeful and creative...Medicine in NYC and author of the book “The Salmon Leap for PhDs: Swimming upstream a career from academia...