...While this approach has been proven feasible, it leaves little room for additional regulatory elements....aureus Cas9 (SaCas9, 1053 amino acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels... indels at a similar efficiency to SpCas9, leading the group to focus their efforts on SaCas9 characterization...Kemp ML, Bao G (2015) Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human cells using compact... (2010) The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71....2020.03.023 Additional Resources on the Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR...

...Jo, Brook, and Angela are co-leading "Making a Greater Impact Using Clear and Accessible Scientific Writing...12 pm - 2 pm. If you want to learn about, and practice, principles of clear, accessible writing through...about, attending sessions, visiting posters, and leading a professional development workshop. Specifically...

...Additional Addgene Podcasts Learn How You Can Contribute to the Addgene Blog Learn about Addgene Plasmid Donations...that will help accelerate his research. Listen to learn more about Dr. Kwapata’s current work and thoughts...us and Seeding Labs for connecting us with him. Learn more about our recent collaboration with Seeding... the Addgene Podcast.     Additional Resources Learn about Seeding Lab's Work with Lilongwe Univeristy...

...on protein therapeutics at Novartis - listen to learn more about Crystal’s exciting career path, to gain...Crystal developed her management skills - training, learning from peers, and being receptive to those who report...science that Crystal works on at Novartis, how she's learned to prioritize projects, and how she sets metrics...thanks to our guest, Crystal Shih. Crystal Shih leads project and scientific teams at the Novartis Institutes...resources in our Mentoring for Scientists Series Learn about Turning Your PhD into a Career with Joanne...

...with and without targeted nucleases). AAV genome editing without a nuclease Although the mechanism of...While the biology behind this AAV effect remains unclear, it is likely not a coincidence that the AAV genome...cells. AAV genome editing combined with targeted nucleases In an effort to combine the respective advantages...introduction of a double strand break by using a targeted nuclease. Asuri et al, for example, used zinc fingers to... and David V. Schaffer. "Combining engineered nucleases with adeno-associated viral vectors for therapeutic...

...., 2012, Bates and Power., 2019): rapid blood clearance, which is useful for imaging applications; better...small size (Bates and Power., 2019). Their rapid clearance from blood can be a drawback for therapeutic applications...of the CAR facilitates signal transduction and release of cytotoxic granules from the T cell. When a CAR...5883 . https://doi.org/10.1073/pnas.85.16.5879  Leavy O (2016) The birth of monoclonal antibodies. Nat...anti-HA scFv-based probe for fluorescent imaging. Learn more about antibodies  Read about another reagent...antibodies, the Developmental Studies Hybridoma Bank Learn more about the SunTag system which uses an scFv ...

...controls, or mixing clear solutions—public speaking is skill that any scientist can learn.  Any time you give...audience something to think about, argue with, or learn from. Some of the best talks I have seen by early...and how the slides complement the spoken message. Learn from their level of explanation in presenting data... science picture. Scientists especially like to learn new things, so rehash common knowledge as little...asked. Reflect on the questions and come up with clear and concise answers. This preparation will help ...

...Daniel Nathans. "Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae... and cars become more technologically advanced leaving earlier versions obsolete, the techniques we use...as their modern equivalents and see what we can learn from their evolution. Southern blotting and quantitative...detection. This increased sequencing speed ultimately leading to what are generally considered first generation...PMCID: PMC4411503. Resources on the Addgene Blog Learn More about Restriction Cloning Get Tips on Analyzing...

...post was contributed by Damien Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc....usually turn into “win-lose” negotiations. This leads to poor outcomes; not enough funds to get started...reduced. Negotiating is easier when both parties are clear on the goals and the resources. Instead of creating...started a company around his own research. He was clear on his science, and found a solution. However extreme...creating and negotiating budgets and invite you to leave a comment below Happy Sciencing, Damien   Damien...Resources Find other career posts on the Addgene Blog! Learn more about Experimental Designs Consulting at their...

...a given section of brain by at least an order of magnitude. It is clear that further refinements of the...inversion of the construct. Sanes and Lichtman used pronuclear injection to insert these Brainbow constructs...effect of multiple, independent recombination events leads to a rainbow of colors. With three copies of the...observed in a given area. In 2013, Dawen Cai, et al. released a refinement of the Brainbow technology, Brainbow...These constructs are stably maintained through at least six generations. In addition to the neuronal pThy1...

[email protected] to learn more. A great submission includes: Images that clearly show viral spread and...reports presented by the submitting scientists, learn more about the AAV Data Hub, and hear the incredible...cellular tropism* (min. 300 dpi, PNG format) At least two replicates of your experiment* Functional data...serotype An abstract that summarizes your experiment clearly and concisely* *required for a DOI. Reports that...

...Blog Learn How We Raised Money for Rare Disease at the Superhero 5k Find New Cancer Resources Learn About...wrote in her application, “Cervical Cancer is the leading cause of cancer related morbidity and death among...researchers who need them the most. Check back here to learn more about future awardees and their inspiring research...Resources on Addgene.org Check out our Cancer Pages Learn about Global Plasmid Distribution ...

...E. coli and other bacterial species. Read on to learn about the tools available for bacteria and some ... and the wild-type sequence persists, Cas9 will cleave the gene of interest, inducing a lethal DSB.   ...systems adapted for eukaryotes in that Cas9-mediated cleavage induces HDR to directly modify a given gene. CRISPR-tranposons...tightly controlled, Tet responsive promoters to avoid leaky activation of the system. Mobile CRISPRi for Gram-negative...VA, Marraffini LA (2014) Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials. Nat...antimicrobials using efficiently delivered RNA-guided nucleases. Nat Biotechnol 32:1141–1145 . https://doi.org... Blog Check out our CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing...

... trainer” programs.  Please join us for the pd|hub Workshop Report Public Release Event scheduled for ...student or postdoc ends up in a lab where they can learn the plus skills needed to succeed on the career ...project and the findings were captured in this just released report “Enhancing Dissemination of Evidence-Based...for March 4th from 2:30-4:00 pm ET.  You can learn more about the report's recommendations and join with...

...world After learning about using sound to visualize bacteria in the lab, I was excited to learn about creative...Fluorescent Protein Resources and get started!  I learned a lot about creative ways to do science communication...communication and outreach - if you'd like to learn more about Science communication check out these SciComm...CRISPR Methods for Bacterial Genome Engineering Learn about Science Communication using Twitter  Resources...

...technology, the ability to direct enzymes – from nucleases to transcription factors – to specific sequences...a cell. Thanks to the work of Huimin Zhao and colleagues, we now have just the tool…designer PUF proteins...possibilities with PUF proteins The tool she and her colleagues led by Zhao described and demonstrated in a report...notes that for reasons that are not yet entirely clear some PUF proteins lose their solubility. Her lab...This isn’t the final version,” she said. “If we learn how to make the library even better, we would be...

...the sample, either by chemical treatment or photobleaching or by stripping out the antibodies, then start... (magenta), anti-AnkyrinG (green), and Hoechst nuclear stain (cyan). Panels C1-C3 show magnified detail... et al., 2021) for more guidelines for creating clear and informative figures. Pro-tip! If you find yourself...antibody, it should get completely washed away and leave no signal. If you see lots of extra signal in these...have questions or any favorite tips and tricks, leave a comment below. Happy imaging! References and ...Wissink, E. M., & Weissgerber, T. L. (2021). Creating clear and informative image-based figures for scientific...

...directly with one another – not just with the team leader. There is no way a manager can be efficient without...Harvard Medical School researcher Connie Cepko to learn about her management style.   Seeking feedback frequently...feedback and so receives this valuable input from at least some employees. Watch out for "information allies...feedback, but shouldn't be the only way.   From a colleague: MBWA "Manage By Walking Around" – don't hide ...Method. P = demonstrate patience, pause; A = ask at least one question to clarify the situation; C = confirm...

...continuity - two lead organizers on each program, one junior and one senior.  To learn more about the STIN's...generation of science communicators. In this chapter we learn about some of their events - science seminars, science...achieving success. Chapter 9: Science by the Pint - We learn why a "Science by the Pint" event is the best way...about starting your own SciComm organization and to learn about some of SITN's other initiatives, email sitnboston...

...productive relationships. I will be sharing what I’ve learned along the way in this ongoing series of blog posts...Harvard Medical School researcher Connie Cepko to learn about her mentoring style. Top 7 questions scientists...well as) their own.” “Someone to act as your ‘cheerleader’ – inspire you.” “Also a mentor can give you ...reach your goals Listens well and communicates clearly and openly Gives honest feedback without ever being... of, and to respect my students as developing colleagues even as I reinforce the necessary power dynamic...