...may provide a method for doing just that. While challenges remain in the delivery of these agents, CRISPR...resistance using multiple approaches. Major restaurant chains including Chipotle and Panera Bread have pledged...remain to help recolonize the niche, reducing the chance of an opportunistic infection like for organisms...quinolone resistance, mediated by a single base pair change in DNA gyrase, CRISPR phagemids were cytotoxic .... PubMed Central: PMCID: PMC4317352. 3. Beisel, Chase L., Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR...

...developing novel technologies to analyze molecular mechanisms of genome functions such as epigenetic regulation...epigenetic regulation. To elucidate molecular mechanisms of regulation of genome functions, we need to...will markedly accelerate research of regulation mechanisms of genome functions in the hope of developing... which is very disappointing. I hope I can help change the situation in the future. Addgene: What advice... they may be able to achieve something that may change the rules of the game. In this regard, I believe...

... Protocol Exchange Website: http://www.nature.com/protocolexchange/ Nature Protocol Exchange (NPE) is ...your favorite protocols for future reference. The exchange currently boasts over 2,500 protocols covering...life science labs and offers the service free of charge to scientists by allowing companies to pay publish...

...buy that charming fixer-upper house you have always admired or volunteer to lead a charitable cause.  ... your work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life... spouse won’t be at every school performance, chaperone every field trip or cheer at every soccer game...your plan to live a good life may be the most challenging assay you will ever have the opportunity to develop...

...Resistance can be intrinsic, meaning a basic characteristic that naturally occurs in the bacterium, and...another bacterium died for instance). These three mechanisms of acquired drug resistance are referred to as...competition drives microbes to develop survival mechanisms. Some microbes even make their own antibiotic...killed), thus increasing the initial microbe’s chances of survival. This was the whole basis of how penicillin...targets (ones that are less likely to mutate or change in order for the bacterium to survive). Check out...

...Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins such ...anti-FP antibodies. GFP emits light in only a single channel. In multi-color imaging experiments, the second...making them difficult to use in a multicolor/multichannel imaging set-up. In contrast, smFP tags can be...University Press, New York. pp 719. 3.Rizzo, Mark A., Michael W. Davidson, and David W. Piston. "Fluorescent ...4. Shaner, Nathan C., George H. Patterson, and Michael W. Davidson. "Advances in fluorescent protein technology...

...methods measure different characteristics of the solution. These characteristics give us information on ...temperature parameters. For example, we found that changing the annealing temperature of the PCR from 60° ...al. "Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated...PMCID: PMC3216358. 4. Lock, Martin, et al. "Characterization of a recombinant adeno-associated virus type...

...better prepare for your career, and increase your chances of landing that next coveted job! Professional...development (or lack thereof) One of the biggest challenges facing students today is that many graduate and...your credentials to HR, greatly increasing your chances of getting a phone interview! If you don’t have...touch and reach back out when you are looking to change positions in the future. Anyone looking for work... Michael G. Lemieux...

...in contrast, engages existing gene regulatory mechanisms to block activity and does not work easily in...into genetic compensation, a previously unknown mechanism of gene regulation (El-Brolosy et al., 2019). ...fluorescently labeled, allowing researchers to observe changes to these tissues under different experimental conditions...time and space, and was also used to precisely characterize developmental defects in the one-eyed pinhead...2016. 219-244. PubMed PMID: 27443928. Harland, Richard M. "A new view of embryo development and regeneration...

...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...So, think beyond Drosophila melanogaster or Saccharomyces cerevisiae (both fine research organisms) -  ...social bonds that they form including genetic mechanisms that control who a female wants to mate with ... first? They are starting by seeing if they can change long legged lizards into short legged lizards.   ...

...counter-intuitive, but I find office work more mentally challenging than lab work at times.” Sometimes a desk job...experience will be a little different. Some of these changes you may enjoy immediately and some of them may ... just take some getting used to. Just remember: change is good. You can experience a real onboarding...to non-scientists can be a fun and gratifying challenge. You’re also getting the opportunity to learn ...eventually. If you approach that transition dreading the change, the adjustment period will be that much longer...

...gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb...the group to focus their efforts on SaCas9 characterization for in vivo studies. One of the pitfalls of...genes, they saw indel formation and phenotypic changes as early as 1 week post-injection. Livers from ...jejuni (CjCas9) is the smallest Cas9 ortholog characterized to date. Kim et al. successfully used CjCas9...transgenes might seem daunting, it’s actually a challenge that the field has overcome with split AAVs. In...

...When you think of Addgene, chances are software development is not one of the things that comes to mind...the flexibility to adapt and easily respond to changing needs of the teams at Addgene. Another advantage...to bake a cake. Instead, you go to a store and purchase flour. However, buying a bag of flour is not the...updated in several years) or they make significant changes that make it incompatible with our software. When...Python 2.7 to Python 3.5. There are significant changes to the language and it took us a total of two years...

...and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and technologies...proteins that could cut RNA or DNA via a CRISPR mechanism. The weird black pellet's DNA yielded a new CRISPR...Prime editing — the ability to now make any small changes, including base edits, insertions, and deletions...right integrase for this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns...be independent of gene repair.) This lowers the chance of the insert being “dropped” in the wrong place...

...Addgene’s YouTube channel.   Figure 4: Global viewership of the Addgene YouTube channel, 2023. Views from...protocols. In 2013, we launched the Addgene YouTube channel to add a strong visual component to our educational...is achieved through interconnected incremental changes. It was only after creating a plasmid repository...learned about the importance of smaller, systemic changes, like using alt-text, captions, and inclusive design...

...inverted vectors expressing light inducible ion channels (plasmid # 20298 for example) have revolutionized...FLEx vectors expressing the light activatable ion channel under the control of Cre recombinase are used in... M. Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range...Sadowski. 1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently...PMC41617. 7. Metzger, D, J Clifford, H Chiba, and P Chambon. 1995. “Conditional Site-Specific Recombination...

...Enzymatic Domains Guide RNA Target PAM (PFS) Cleavage Mechanism Cas9 HNH, RuvC TracrRNA, crRNA DNA 5' NGG... Leptotrichia shahii (LshCas13a) to begin to characterize this new effector (Abudayyeh et al., 2016). ...and cleavage through a series of conformational changes in the Cas13a molecule. Like Cas9, Cas13a tolerates... plant cells (Abudayyeh et al., 2017). They characterized Cas13a from Leptotrichia wadei (LwaCas13a), ..., Sergey, et al. "Discovery and functional characterization of diverse class 2 CRISPR-Cas systems." Molecular...

...to interfere with the function of resident ER chaperones and may impact cellular function or possibly ...For example, the formation of inappropriate inter-chain disulfide bonds between FPs blocks Golgi complex-localized...protein activity. We encourage investigators to characterize FP-fusion proteins functionally relative to ...moxFPs. The inherent brightness and spectral characteristics of the moxFPs are comparable to the non-optimized...postdoctoral fellow at the University of North Carolina at Chapel Hill. Under the co-mentorship Blossom Damania and...

...found it easy to drop into a chair and chat with me remotely via an audio chat (Fig. 4).  Learn: This works...small operation, so sometimes it can be a big challenge. But ask any filmmaker or videographer and they...you can usually see a director sitting in their chair not watching their actors live on set, but peering...

...sensor relies on a change in localization to the cell membrane and not a direct change in the fluorescent...better response kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and...   Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium... B) LiLac responses are highly resistant to pH changes. Image adapted from Koveal et al. 2022, under CC-BY...