...many of these studies report correlations without clearly testing whether a certain methylation state causes... recognized by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic... or throughout the genome – could be affected, leading to unpredicted gene expression changes (Sapozhnikov...simpler approach - dCas9 blocks DNMT1 methylation, leaving the target CpG unmethylated. Adapted from Sapozhnikov...Sapozhnikov and Szyf 2021.   Let me leave you with a few practical considerations: Design your gRNA...an unmethylated CpG to behave naturally in the nuclear environment. One good system to balance these needs...

...and lentiviral envelope plasmids are listed here. Please note that both lentiviruses and gamma-retroviruses...express siRNAs of their choice in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi! Cre-Lox... gene knockdown. Vector provided by the Qi lab. Learn more about CRISPR plasmids for transcriptional inhibition...mammalian genome that can reveal how genetic alterations lead to changes in phenotype. The plasmid comes with ...We are constantly improving our CRISPR reagents. Please check www.genome-engineering.org for the most up-to-date...information. Check our CRISPR guide and blog to learn more about CRISPR and the most recent developments...

...Addgene to accelerate their research. Read on to learn how these researchers plan to use plasmids from ...Sciences of NOVA University in Lisbon, Portugal. She leads the Glycoimmunology group based at the Research ...mild or exacerbated inflammatory response which leads to nearly 25% child mortality. Since glycans play...human disease change Arg to another amino acid. Clearly, the development of a pharmacological intervention...binds to and inhibits a potassium channel, causing release of acetylcholine. Given its unique molecular biology...BeHeard Award winners! Paula Videira is the team leader of the Glycoimmunology group at UCIBIO, FCT-UNL...

...We started with learning the head is connected to the neck. Shortly after, we learned about organs and...connectivity? One of the early lessons many of us learned in biology is that the body’s architecture and ...terminals (Lafon 2005). Therefore, once RABV is released from an infected cell, it is only taken up by ...genome). Nascent G protein-coated RABV is then released from the starter cell and transmits retrogradely...mediated by other viral proteins), but the newly released RABV cannot infect any subsequent cells due to...

...the future of the bioeconomy it's less clear, but the way I'd leave it is mostly to the imagination. The...collaboration this way: "Addgene is one of the leaders in distributing plasmids amongst the academic community...Addgene: And the BIL gates collection? Endy: The Boolean Integrase Logic (BIL) gates collection of parts...basic logical functions, which we know and love. Boolean is in reference to George Boole who in 1854 was...incredible hidden story here. The parts we are releasing simply plug into this existing economic activity...

...CRISPR plasmids containing a nuclease into three functional groups: nucleases, modulators, and chimeric ...chimeric effectors. Nucleases generate a classic double-stranded break, while modulators regulate transcription...function, from 2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain the most popular...educational resources help scientists navigate that learning curve. As of 2023, our CRISPR guide has been viewed...around the impact of science, but our data makes it clear that it’s also a story of community, cooperation...

...in his laboratory. Learn more here. Additional resources on the Addgene blog Learn more about plasmid...across regions, with Europe most impacted and China least impacted. The inferred weekend effect is, in retrospect...date range is, they could be the most impacted or least impacted. These results highlight the instability...candidate at MIT. His research uses applied machine learning and statistical methods for fundamental scientific...interested in using selection, evolution, and machine learning to solve difficult problems in chemistry and chemical...

...there has less chance of contamination with RNase leading to degradation that can set you back days. Working...(e.g. face mask, and a dust free and specially cleaned work bench). Once the pups are born It takes roughly... variety of phenomena, it’s possible that we’ll learn much more from our favourite mammalian model organisms...off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1 (2014): ...Daisuke, et al. "Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9...Engineering with CRISPR Validate Your Genome Edit Learn about Non Homologous End Joining and Homology Directed...

...personal time. Someone else can clean your house, pick up your dry cleaning, cook your dinner and deliver...fixer-upper house you have always admired or volunteer to lead a charitable cause.  While I can’t make it “all ...and tactics that have worked for my friends, my colleagues and me. To that end, here are my personal 10 ...happen. Be realistic and: Your co-workers will learn to work within these bounds You can work later (...much more confident about their safety when they leave home which happens very, very fast! I will never...

...our CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips...sequences Cas9 likely goes through multiple rounds of cleavage at the target site until either the PAM site or...arrested too long. Prolonged cell cycle arrest can lead to apoptosis in as little as 10 hours, so read up...Cas break for HR is to fuse an HR protein to the nuclease. Early HR factors fused to Cas9 have been shown... screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to...

...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...

...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...

...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...

...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...

...., 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 ...

...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...