...heterodimer in response to blue light, which will ultimately co-localize the VP64 transactivator with a dCas9... innovative ways. The principle of protein multimerization is seen in many important biological contexts...copies of a protein to a target polypeptide. Multimerization in the SunTag system occurs via antibody-peptide...single-molecule reporter of intracellular processes. Multimerization via the SunTag system also shows strong upregulation...specified cellular processes or be observed at a later time via DNA sequencing or other readout. Cellular memory...large changes in binding affinity with light stimulation. Indeed oLID showed only a two-fold change in...50 fold change in binding affinity with light stimulation2. The Kuhlman lab created two improved Light ...

...they have a relatively short generation time for mammals - the time between being born and giving birth -...maintained in the lab, and have short generation times that make it easy to study the effects of genetic...simultaneously observe several generations of mice at a time. Because mice are more genetically and physiologically...potential for therapeutic drug discovery as it is estimated that 75% of disease causing genes in humans have...other model systems but can be quite difficult and time consuming. GAL4/UAS was first described in 1993 ... a set of modular vectors that can be used to optimally control gene expression in specific cells of the...groups, easily bred, and produce 50-300 eggs at a time. Zebrafish embryos are also laid and fertilized ...

...editing accuracy by preventing re-editing and (2) optimizing “cut-to-mutation distance”. Furthermore (3), ... strategy that was discussed in the field at the time but had never been systematically analyzed: introducing...find the right one Step 2 – Edit the cells a second time using a repair template that corrects the blocking...CRISPR Resources on Addgene's CRISPR Guide Page 2. Optimizing "cut-to-mutation distance"  Even though we had...mutation distance and editing efficiency.   3. Optimizing "cut-to-mutation distance" for homo or heterozygous...that for generating a homozygous HDR event with optimal efficiency a guide RNA targeting a cut <10 bp from... other on each target site.   Figure 3: Optimal cut site to mutation distances for heterozygous...

...cryo-EM grid freezing Engineered and evolved optimizations for prime editing CHARM: A compact epigenetic...requiring a lower concentration of sample — nearly ten times less protein. While the addition of LEAs may bias...2024.02.06.579238.    Engineered and evolved optimizations for prime editing By Emily P. Bentley The David...Liu Lab has made two recent deposits focused on optimizing prime editing systems for specific applications...bp, PASSIGE can integrate cargoes over a hundred times that size (demonstrated up to 10.5 kb) by relying...evolves and engineers Bxb1 recombinase variants to optimize PASSIGE efficiency (Pandey et al., 2024). The ...four-fold improvement over the wild-type enzyme. Find optimized PASSIGE plasmids here!     Figure 3: Schematic...

...These are stormy times. It feels like we’re being buffeted by wind and waves and sometimes wonder if we can...Thanks to everyone for being awesome during a tough time. We're all going to get through this and have one....” And we sent the Addgenies home.  At the same time, as Addgene started dealing with this new reality...available to industry requestors as well. This is a time when we need as much open science sharing as we ...’re all doing amazing work during this difficult time. One function that rose to the challenge was our...with labs worldwide closing for unknown amounts of time, we were facing a financial crisis. As a nonprofit...keep new projects on track to succeed at the same time. I began approaching science funding organizations...

...of the time in a test set of 173 known synthetic and natural genes.  In designing and optimizing their...how Aditya Kunjapur, a MIT graduate student at the time, got involved. Along with Philipp Pfingstag, a visiting...visiting graduate student from TU München at the time, the team built an algorithm that predicts whether...codon usage biases. But if a gene’s sequence was “optimized” so that it more closely resembles that of the...genes where there are instances of rare codons, sometimes intentionally modulating expression in ways we...-of-origin of engineered DNA This isn’t the only time researchers have used Addgene as a large data set... gene are essential and complementary tools in a time where biological engineering and synthetic biology...

...interest, and stimulate or manipulate responses through neurological means like electric stimulation, drug administration...response to both of these stimulations. The best part? The chemical stimulation was sucrose…..sounds like... The fibers they ended up generating were 10-15 times more flexible than the brain fibers and importantly...microcontroller, NeuroStack coordinates optical stimulation of its associated fibers across multiple channels...authors first tested that the system could induce stimulation in target tissues of a live animal. They started... to the brain tissue. They performed optical stimulation using NeuroStack and the fibers as previously...

...choose could be based on your downstream application, time, and instrument availability. UV absorbance Using...ug/ml pure dsDNA. Use the following formula to estimate your DNA: Concentration (ug/ml) = A260 reading...quantify your DNA prep. Though this method takes more time to set up in the lab, you might not have to do the...electrophoresis, you only need 1-2 ul of sample and the run time is just a few minutes per sample. During the run...larger fragments. These fragments are quantified over time using a fluorescent dye that intercalates into the...electrophoresis which shows fluorescence units over time. Smaller fragments migrate more quickly than larger...that can be quantified at 595 nm. This method is time consuming and has low sensitivity so it’s not used...

...that get smaller in size as a function of digestion time. For phage DNA that is refractory to enzymatic digestion...be selected for by manipulating the nebulization time and pressure. Nebulization units, which can be purchased...fragments falling between 1 and 3 kb results in optimal likelihood of capturing a whole RBP gene without...80°C) followed by 5 min at 37°C (each repeated 5 times) works well. Leaving the membrane overnight in the...to enhance folding of the recombinant proteins. Optimal salt concentration will vary with each protein,...at room temperature as opposed to overnight at optimal growth temperature. If you’re lucky enough to have...the membrane to probing with cells in order to optimize this portion of the assay before attempting it...

... in live cells. The fluorescent timer emits blue light around the time of synthesis and, after a few hours...Fluorescent protein system for C. elegans Fluorescent timers for studying cell cycle length The CRISPR corner...pattern of hippocampal adult-born granule cells over time that were not previously possible.   Bensussen ...-wrmScarlet is a new split red FP that is three times brighter in worms than split-sfCherry3. It was created...cell cycle length with a histone-fused fluorescent timer By Kate Harten DeMaio Studies of the cell cycle ...Yale University fused a color-changing fluorescent timer (FT) protein to the histone H2B that allows for  ... Get the plasmids at Addgene Sc++ is a codon-optimized, engineered Streptococcus canis Cas9 nuclease ...

...transfected and harvested your lentivirus. Now it’s time to move ahead with your infection and make your ... of virus you actually have in your prep. Taking time to properly titer your virus will not only ensure... the best possible way, but it may also save you time in the long run. Read on for an overview of the ... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...assay, titering via measurement of viral RNA can overestimate the amount of infectious virus in a prep as ...physical titer, there is a risk that they may underestimate viral titer as these assays cannot distinguish...integration events, reducing the possibility of underestimating the viral titer. The main drawback of this ...

...industry postdocs have a set time limit. At AstraZeneca, this is three years. This time limit can feel like a...might have less time to explore scientifically since you can’t afford to spend too much time working on something...Pros and cons: The academic vs. industry postdoc  Time: Academic postdocs can be long—5 years or more, ... Also, if you work in industry for any length of time, you’ll know more people in industry, which means...numbers of publications, which may take a longer time to acquire. Learning about drug discovery: Unless...as AstraZeneca, will hire their postdocs for full-time positions should a position open up. Other companies...

...small amount of DNA can be copied many many times in a short time period. PCR was invented in the early 1980s...denaturation, annealing and extension is repeated 25-35 times to exponentially replicate the target DNA of interest...which would slow or stop PCR amplification. Other times, the PCR reaction may not be 100% efficient and ...polymerase chain reaction (qPCR) also known as real time PCR is a PCR technique used for measuring a starting...require a reference or standard curve, which can be time consuming and challenging to get right. ddPCR uses...an extremely useful PCR method that can help save time and effort in the laboratory.  There are two main... different pathogens in one specimen saving both time and effort. Scientists can also utilize multiplex...

... anaerobic conditions. Maturation Time: Maturation time is the time it takes the FP to correctly fold ...Dreiklang). Fluorescent Timers (FT): These proteins change their color over time. Therefore, these can ...can be used as “timers” for cellular processes following their activation. The four main FTs are called ...and DsRed ~10hours. Temperature: FPs maturation times and fluorescent intensity can be affected by the...temperature. For instance, enhanced GFP (EGFP) was optimized for 37°C, and is therefore most suited for mammalian...

...are some tips and tricks you can use to obtain optimum transformation efficiency. It can take 6-8 weeks...pretty important to get this process right the first time. I’ll lay out some guidelines here. A detailed transformation...transform early, your plants will still have plenty of time to produce seeds after you expose the flowers to...you are transforming more than one construct at a time. For detailed protocols, volumes, spin speeds, ...agrobacterium: Getting a transformant takes a long time. Make sure the agrobacterial culture you are growing...cells in your culture will no longer be viable. The time this will take depends on the volume of culture ...inoculated from a colony on a plate will reach an optimal density in 18-24 hours. I then add 0.5 mL of this...

... for structural studies of E3 ligase cereblon Optimized mScarlet-based tags for live imaging in C. elegans.../10.1038/s41467-024-52871-9. PMID 39406745. Optimizing mScarlet tags for live imaging in C. elegans ...for mScarlet3 and mScarlet-I3 that were codon-optimized for C. elegans and included artificial introns...mScarlet3 was 50% brighter than the original worm-optimized wrmScarlet. mScarlet-I3, while initially dimmer... et al., 2024 under CC-BY license.  These optimized fluorescent proteins will be a useful addition...efficiencies of up to about 9% of colonies (102 to 104 times higher than traditional homologous recombination...mechanical forces with PILATeS By Emily P. Bentley Time to get flexible! Alex Dunn’s lab and collaborators...

...loxP sites (For details see Cre-lox blog post). Optimizing FLP FRT through smart engineering Different FLP...first version of FLP discovered has a temperature optimum of 30 °C and is therefore inefficient in mammalian...identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998)....less efficient than Cre recombinase. FLPe codon optimization resulted in FLPO, which shows similar efficiency...the recombination should only occur at a specific time point? To allow temporal control, ligand regulated...and its synapse should be visualized at the same time from two different plasmids this approach becomes...rearranges these synapses in the living brain over time (Villa et al. 2016). Combing FLP and Cre for additional...

... the better (Nat. Biotechnol., 2014). Given the timing, it shouldn’t be a surprise that the CRISPR system...software to make sgRNA design accessible to all. At the time I thought there was room for improvement and a year...iterative design; it is caused by not having the time to assess public response; and, it is the stage ...winner, is there something that we can invest our time into learning and extract the most value? This is...and not general opinion. I could spend a lot of time running through each tool in detail, but that is... genomes of any size and are only limited by the time it takes authors to add a genome to the tool’s repertoire...those who are entering sgRNA design for the first time, it is best to choose a tool based on immediate ...

...one of the earlier epicenters of the outbreak ultimately flattening the curve, it has become evident that... reagents and equipment and the long turnaround times, have led researchers to turn to other technologies... Sensitivity:10-100 copies of RNA per microliter Time: ~40 minutes Equipment required: 37°C heat block... developed so far.  A similar method has been optimized to detect the SARS-CoV-2 virus directly using ...milliliter can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath and...microliter of RNA targets in a 40-minute incubation  Time: ~90 minutes Equipment needed: 37°C heat block, ...CREST) Sensitivity: 10 copies of RNA per microliter Time: Not mentioned in the paper, likely approximately...

...from paper to an electronic lab notebook requires time and knowledge. In a recent article, we translated...your research can be patented? I.e. is automated time stamping (detailed records on who did what and when... when) important to you? Some ELNs record the times and dates of your actions. These records can take ...about every action performed in the lab (user, date, time, action) are required for regulatory purposes in...them.  Conclusion Researchers spend 40% of their time on tedious administrative tasks that drain resources...resources from the actual research process. It does take time and effort to implement an ELN, but on the long ... organized within an ELN already. Investing some time in deploying a software solution that makes it much...