... any preventive measure designed to help keep you from getting sick can be helpful in preventing the spread... on preventing the spread of antibiotic resistant bacteria, check out this resource on preventing antibiotic...because ‘antibiotic’ merely means that the drug prevents the growth of a living organism. As for multi-...standpoint, we can all work together to try and prevent outbreaks of drug resistant bacteria within communities...standards. Handwashing is an essential component of preventing infection, as is cleaning and disinfecting frequently...

... and the promoter to prevent leaky expression of this resistance. Unlike previous systems, pUCXKT does...can do so in a very specific, precise manner. Previously, scientists have been successful in regulating...enzymes (e.g. nucleases, recombinases) which irreversibly excise or invert a specific chunk of DNA. However...deposited a set of 11 phage integrases which act irreversibly on their cognate attB/attP sites with no cross-talk...subcellular localization in mammalian cell culture and reversible control of signalling pathways. These LIDs are... powerful tools which allow you to easily and reversibly switch off and on your favourite signalling pathways...the MCS has several options) and a pUCX-specific reverse primer containing the missing codons from the antibiotic...

... have taken researchers to the threshold of a revolution in modern neuroscience, an era in which technologies...antisense orientation with respect to the promoter to prevent its expression, and is flanked (“floxed”) by two...electroencephalography evidences of arousal in mice previously subjected to steady-state general anesthesia ...behaviors. The high spatio-temporal precision and reversible modulation of FLEx vectors combined with the ...functions, and molecular mechanisms." Chemical reviews 114.1 (2013): 126-163. PubMed PMID: 24364740. PubMed...development and application of optogenetics." Annual review of neuroscience 34 (2011). PubMed PMID: 21692661...of cre-loxp site-specific recombination." Annual review of biophysics and biomolecular structure 30.1 (...

...plant hormones called auxins to conditionally and reversibly degrade proteins of interest fused to a degron...2009, Natsume et al., 2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method to tag endogenous... be used to engineer specific sites that were previously difficult. Find SpCas9 and Target-AID-NG plasmids...promoter and an enhancer. What makes LADL unique from previous iterations is the addition of CRY2, another Arabidoposis...hours of blue light induction (6-fold faster than previously developed systems). The LADL system promises ...it’s briefer time scale and its inducible and reversible nature. Find the LADL plasmids at Addgene Kim...

...Turn it on? Turn it off? Turn it down, but not forever? You don’t need to resort to the force to control...biological tools are abundant! In this post, we will review tried-and-true protein expression control tools...the options together. Turn it off Turn it off forever If you want to turn off protein expression, you...for transient knockouts!  Turn it on Turn it on forever Permanently increasing or turning on expression...amounts of tetracycline. Tetracycline regulation is reversible (when the drug is removed expression goes back...doesn’t require this level of control, one of the previous options may be better suited for the job. Whether...

... issue is reviewed yearly at a best practices meeting, where the QC team updates and reviews our sequencing...caught prior to distribution. Catching these errors prevents future experimental issues, saving time and money...encountered in plasmids deposited to Addgene. This review process also highlights other QC topics that may...team suspects an assembly issue, they manually revisit the raw NGS reads and reassemble using a program... issue, they will contact the depositing lab to review the discrepancies. Often, the identified issues...

...that relies on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing guide RNA (pegRNA...binding site (PBS) complementary to the genome and a reverse transcriptase (RT) template including the desired...Anzalone et al., 2019). Test different lengths of reverse transcriptase template, starting with about 10–...optimizing your pegRNA primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com...Edit the PAM along with your intended edit. This prevents the Cas9 nickase from re-binding and nicking the... al., 2019). Figure 4: Editing the PAM prevents the prime editor from re-engaging with DNA it ...

...is published. Plasmids can be linked to a peer-reviewed manuscript OR a preprint. In the case of a preprint...information once the manuscript is published in a peer-reviewed journal. To start this process, simply enter your...Step 5: Verification On this page you will see a preview of the final plasmid page that will appear on Addgene...once your plasmids are available online. Please review all of the information on this page to make sure...easily update the information by navigating to the previous pages using the links at the top of the page. .... Figure 11: A completed plasmid page for review before submission. Once you are satisfied with the plasmid...backbone, you can copy some information from a previous plasmid in your deposit by selecting the plasmid...

...one mistake can be enough. Reversal drives are designed to overwrite a previous gene drive and thereby undo...by the earlier intervention2. While an initial reversal drive cannot restore the exact original sequence... sgRNAs and possibly cas9 gene). An immunizing reversal drive is a variant that also spreads through the...should consider building an appropriate immunizing reversal drive at the same time to mitigate the potential...M. RNA-guided gene drives can efficiently and reversibly bias inheritance in wild yeast. bioRxiv 013896...

...fill the prop with something of value. They are reviewers of content. On the other hand, the goals of the...defined by their trust in the piñata builders - the reviewers. After some physical exertion on the part of the... the value of each item changes from what the reviewers so painstakingly assessed to something a bit more...”, it is the disconnect arising between those reviewing the contents of the piñata and the audience blindly...software development has caught up with it. The peer review process has been challenged with many closely spaced...variations on a theme without many resources to review them. Collectively, the pool of CRISPR software...view and get a sense of which sgRNA designs have previously worked and under what conditions. Such resources...

... to review grant proposals from others. Dr. Cook was asked to serve on review panels and to review education...profession? I asked Deborah Cook, Ph.D., GPC, previously a college biology professor, how she became a...development? Volunteer as a grant writer and/or grant reviewer with a local nonprofit that aligns with your passion...Profession. Whether you’re transitioning from a previous career or you’re starting your first career, a...

...invert or ‘flip DNA’, is derived from the yeast S. cerevisiae (Gronostajski and Sadowski 1985). The FLP system...recombination events mediated by FLP or Cre are reversible. Whereas the excision of a piece of DNA flanked... within that sequence and should therefore be prevented. The loxP and FRT target sites have been engineered...sites (Schnütgen et al. 2003). Flex vectors revolutionize neuroscience Manipulation of specific cell types...channels (plasmid # 20298 for example) have revolutionized Neuroscience. They are the only means to target...1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to DNA via...

...activating genes that cause differentiation and to reverse HIV viral latency (Bialek et al., 2016). Additionally...2015). These types of CRISPR activation are also reversible: once the environmental trigger leaves, gene ...activated. However, there are several factors that may prevent your gene from being up-regulated. For instance...This is believed to be true given the results of previous RNA-seq experiments (Konermann et al., 2014, Chavez...Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator...10.1016/j.cell.2014.09.029 Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib...

...photostability under two-photon illumination than previous GEVIs. Some constructs are also available as viral...kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and...reduced sensitivity to pH and calcium compared to previous lactate biosensors, and its affinity is better-matched...temperature sensor that significantly outperforms previous designs. B-gTEMP is excitable under blue light...B-gTEMP itself is a significant improvement over previous temperature indicators, it also demonstrates that...Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature Imaging with a Genetically...

...suggest that phosphorylation is far more prevalent than previously appreciated. PhosphoSitePlus, one of the... group. Additionally, these mutations are not reversible, and thus cannot reflect the true state of protein...Extensive genomic sequencing of prokaryotic lineages revealed a curious anomaly amongst a subset of methanogenic...mutation of these residues. Due to the limitations previously mentioned, phosphomimetic mutations on the activation...affects protein function, researchers should try to reverse the effects of pSer-incorporation on enzyme function...

...the continued utility of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need ... removes one of the other loxP site variants, preventing further recombination. Brainbow-1.1 scales up...that transient Cre recombination is required to prevent constant inversion of the construct. Sanes and...their protein sequences are highly homologous, preventing design of antibodies specific to each fluorophore...of delicate axonal and dendritic processes not previously visible with Brainbow-1 and -2. The general structure...connectome. Lichtman JW, Livet J, Sanes JR. Nature Reviews Neuroscience. 2008 Jun;9(6):417-22. doi: 10.1038...

...as a nonprofit under the United States Internal Revenue Code 501(c)(3). That means we were formed to benefit... are able to fulfill our mission by generating revenue through distribution of research materials. Our..." is a bit misleading. Nonprofits can generate revenue (as Addgene does through materials distribution... nonprofits should have some level of positive revenue to build reserves so the company is sustainable...The difference is what nonprofits do with their revenue. A nonprofit organization cannot distribute profits...

... news is that this effect is reversible and the pH shift can be prevented. Keep this information in mind...if you’re receiving samples on dry ice - it may prevent you from seeing some unexpected results. Order... 3). However, 30 seconds was not sufficient to prevent any pH change, as we still observed a slight change... allowed the CO2 to leave the tubes, and would prevent the pH change upon thaw (Fig.4). To test this, ...

...entirely too much work for one person, it would take forever, and the end product would likely be poor quality... delegate the giving of feedback or performance review. You can’t delegate tasks assigned to you and you...outcomes, timelines and deadlines. Agree on a way to review project progress. I recommend putting a regular...approachable mistakes will be made that could have been prevented.  Make sure everyone understands that no question... Aren't You Delegating" in the Harvard Business Review. Another excellent blog on delegation tips and ...

...your vocabulary! A brief review of base editing tools First, let’s briefly review how CRISPR-derived base...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...the pyrimidine bases (C and T). (Check out our previous post on these editors for more details on how ...also called apurinic or apyrimidinic sites and abbreviated as AP sites. To explore this further, the team...bound, keeping the two DNA strands separate and preventing repair based on the complementary strand. This...