...polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific end cleaning enzymes (Aprataxin, Tdp2...Cas9 protein doesn’t immediately release from the broken end after cleavage, which may interfere with loading... the products of accurate repair are easily re-cleaved while indel products aren’t, so repeated cycles...latter products. As noted above, a single cycle of cleavage and accurate repair should take less than an hour...

...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or genes to...published in Nature in December, Konermann and her colleagues in Feng Zhang’s lab describe just such a system...transcription activation processes. Konermann and her colleagues demonstrated the new system by screening for ...in those 15,977 human genes. Weissman and his colleagues demonstrated in a paper in Cell that they could...which guide RNA we use. What we have to do now is learn more about the rules.” He said that there is no ... These libraries are lentiviral in nature, so please ensure that you are equipped and authorized to ...

...review see Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal...expansion of RABV. This improvement resulted in a clearer picture of neuronal connectivity in the mammalian... mapping of the neuronal circuitry involved in learning and memory (Kohara et al., 2014; plasmids at Addgene...particle is taken up on one side of the cell and released again at another side of the cell after intracellular...transport without actually causing infection or leading to transgene expression within the first cell. ...development of more powerful AAV serotypes, and perhaps lead to controlled trans-synaptic spread of AAV vectors...21782903. McFarland, N.R., Lee, J.S., Hyman, B.T., and McLean, P.J. (2009). Comparison of transduction efficiency...

... and ligands vary in their chemical structure, leading to differences in their excitation and emission...peak wavelength can also decrease the rate of photobleaching. Interaction with receptors: Understanding ...the choice of fluorescent dye. For instance, the cleavage of FITC-conjugated protein ligands is unaffected...experimental conditions. Common issues include photobleaching, non-specific binding, and quenching, which... reduce ‘muddying’ of the signal and facilitate clear differentiation between multiple dyes during analysis...states in response to a specific stimuli, which leaves only a small volume of fluorophores in an excited..., M., & Ghods, R. (n.d.). Comparison of the Photobleaching and Photostability Traits of Alexa Fluor 568...

...that other technologies need to be discarded but clearly we are in an era of neuroscience research where... the technique is remarkably straightforward to learn and relatively cheap (compared to many other technologies...Program (NIH-NIDA IRP), with the stated goal of learning behavioral optogenetics in order to implement ... to share some of the tips and tricks and I’ve learned from numerous scientists at NIDA and some of the...region can result in a particular neurotransmitter release or post-synaptic current. Fortunately, if your ...variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin variants and how ...University.  His interests include the role of learning and memory in opioid addiction and science communication...

... into close proximity with the target protein, leading to polyubiquitination of the target protein and...proteasome machinery through binding with cereblon, leading to rapid target degradation (Figure 1). Plasmids...ortho-AP1867) via various chemical linkers. Our current lead dTAG molecule is dTAG-13 due its immediate action...happy to share aliquots of dTAG-13 in the meantime, please address your requests for dTAG molecules and any...terminus is favorable for tagging. However, if it is unclear which terminus should be tagged, the dTAG lentiviral... interested in targeted protein degradation, and lead author on the recent paper describing the dTAG technology...

...she made through this program lead to her current job. “One of my colleagues in the class was like, “Hey...people are,” says Susan. Her passion for insects lead her to start a PhD in entomology at the University...which Susan replied, “What’s a science writer?” Learning about SciComm by doing SciComm Writing had always...Evaluating evidence. Critical thinking. Being able to learn things quickly. Story telling. The ability to know...tips for using social media as a scientist here Learn more about storytelling in science ...

... this cause and helped by expanding this team, leading monthly calls, and planning next steps. April Clyburne-Sherin...future of reproducibility workshops and lessons learned: Since early 2018 the team has grown to include...all levels of the scientific community. Lessons learned Instead of continuously talking about a problem...like to join or contribute to this collaboration, please check out the official website, follow the group...conversations on the history of the group, lessons learned, and to Sonali Roy, Magdalena Julkowska, and Jennifer...on Transferable Skills: External Collaborations Learn about the 2018 Minisymposium on Scientific Reproducibility...

...metabolism. Cancer cells promote oxidative stress leading to the degradation of one key enzyme (among others...built to protect us now seems to attack us by releasing mitogenic growth mediators and improving cancer... When it comes to breast cancer, Hoshinho and colleagues found that after injecting breast cancer cells...process is not fully understood, a team of researchers lead by Takayuki Kogure (Kogure et al., 2011) believes...Addgene blog Read our cancer related blog posts Learn about our cancer collection pages Read more about...resources page  Find plasmids for your research Learn about Addgene's viral service ...

...these molecules are clearly associated with cellular damage and dysfunction, it is unclear what the precise...Transposable elements such as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription...silenced, but dysregulation of these systems can lead to local dsRNA production. In the same vein, genotoxic...barring a loss of mitochondrial integrity that would release them into the cytosol. Fig. 2: Endogenous... of DNA damage, and these structures can in turn lead to genome instability. Thus, it’s important to strike...

...s lab and Viviana Gradinaru’s lab used machine learning to design new channelrhodopsins that are more ...channelrhodopsin that’s successful in vivo. Using machine-learning to overcome channelrhodopsin design challenges... Arnold and Gradinaru Labs developed a machine-learning model to help them construct “designer” channelrhodopsins...and secure a fiber optic cannula to the skull, leaving the brain intact. When the fiber optic light was...Mackey ED, Gradinaru V, Arnold FH (2019) Machine learning-guided channelrhodopsin engineering enables minimally... PHP.eB AAV capsid Read more about optogenetics Learn about the differences between chemogenetics and ...

...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...

...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...

...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...

...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...

...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...

...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...

...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...