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...recombinase systems. Picking Ligands Picking the right ligand is also important for building a chemogenetic...pharmacologically selective effector molecules (PSEMs). Higher potency means lower drug doses, which decreases... CH, Coward P, Degtyarev MY, Lee EK, Kwa AT, Hennighausen L, Bujard H, Fishman GI, Conklin BR (1999) Conditional...

...lucky" and find a great gRNA with a proximal PAM right where you need it, the odds of this occurring are...reliability. Notably, the SpRY variant didn’t display higher off-target effects than SpCas9, a pleasant surprise...stranded cleavage and base editing applications, with high editing capacity in both contexts for NRN PAMs and...

...host) as well as the introduction of the cancer-fighting component, CAR. Clinical trials are underway to...allele specifically. Early-stage trials showed a high success rate in clearing misfolded protein levels...Regarding effectiveness, if the editing frequency isn’t high enough to make a physiological difference, then ...

...endogenous retrovirus Rless glycoprotein (BaEVRless) had high rates of transduction for most cell types they tested...existing CRISPR delivery technologies, Nanoblades had higher rates for editing efficiency and lower rates of...zygotes, which results in albinism in mice. Of the eight mice born from Tyr Nanoblade treated zygotes, five...

.... In the first blog post in this series, I’ll highlight terminology you should be familiar with before...the use of long single-stranded DNA have led to a higher efficiency of homologous recombination for precise...

...understanding of cell biology. We hope that these high-quality, highly specific plasmids, as well as the instructions...flooding the cell with a plasmid containing regions of high homology to the gene with the cut, we trick the ...

...company. This might seem like a no-brainer, but it happens more frequently than you might think. The Addgene... crucial information about a product is printed right on the tube itself! Is the answer to your question...

...something silly like “See there, arginine 53 really is right there, at the end of helix 3…just like we said it...are useful. While a physical model of a protein might represent one aspect of the protein very well, it...while and you will wake up in the middle of the night dreaming about the conformational gymnastics that...

...colonies by (carefully) holding plates up to the light, where they appear as opaque dots (colonies should...the agarose onto the recipient plate at just the right temperature. If the agarose solution is too hot,...depending on the level of endogenous expression. We highly recommend the use of an electronic pipettor with...

...primers.  Extension- The temperature is then raised slightly to Taq polymerase’s ideal temperature (70-75oC...which can be time consuming and challenging to get right. ddPCR uses a water-oil emulsion droplet technology...those stated above. Multiplex PCR is also used in high throughput SNP genotyping, gene deletion analysis...

... of interest thus represents a straightforward means of exerting tight control over Cas9 nuclease activity...different genetic loci within a single cell. One might imagine transfecting cells with both the nuclease-positive...

... Sometimes people don’t know how to start. They might want to use our promoters and terminators, but then...coding sequence to the MoClo system. It’s quite straightforward really and we’ll be producing an instructional...chances of doing something amazing will be equally high as for those who go to work in cancer biology. ...

...to make the most of your time and sequence! You might also enjoy seeing how our quality control process...alignment results. The default setting is “Optimize for Highly similar sequences (megablast)”, which is very fast...nucleotide sequences, proteins often have repeated or highly homologous regions, which by default are ignored...

...a collection of promises to not assert property rights for particular engineered genetic sequences; but...metabolite in a cell. The only way to do that at scale right now is to grow up a bunch of cells, sacrifice them...chemicals, to make more medicine. The parts we have right now are well adapted to work in prokaryotic bacteria...

... off-target effects. Cas9 RNPs are detectable at high levels shortly after transfection, and are quickly...efficiency, while biolistic bombardment, which can have high efficiency, requires specialized equipment, such...microbiology.     References 1. Chen, Sean, et al. "Highly efficient mouse genome editing by CRISPR ribonucleoprotein...

... know the basic principles of IF, we’ll take a slightly deeper dive into those steps and provide you with...commonly used cross-linker for IF. However, you might come across a couple other terms that can cause ...especially if left on for too long or used at too high a concentration.      Figure 2: Examples...

...A) Streamlined VchINT expression constructs. (B) High integration efficiencies achieved with VchINT, at...mini-transposon Transform plasmid(s) into cells and plate overnight Screen colonies phenotypically or by PCR/sequencing...Sternberg SH (2020) CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering...

...trainees’ very way of thinking too. Albert Einstein might have put it best when he stated, “the value of an...pathways (precisely the type of exercise Einstein might have resented!). In 1997, Joseph Provost, then a...like an exciting opportunity for your classroom, I highly recommend you try implementing one. It's likely...

...available Not commercially available Small but Mighty: Advantages of Nanobodies Although antibodies have...target antigen to about 2 nm, which can lead to higher resolution for super-resolution microscopy. Nanobodies... two questions, refer to this table to find the right nanobody for your work. Species Antibody Subclass...

...standard curve for qPCR can be challenging to get right. Here at Addgene, we use a linearized plasmid standard...very few positive droplets. In the image to the right, there is approximately 1 positive copy per microliter...droplet generator to the PCR plate. Maintaining a high droplet count is important for calculating the concentration...

...particular gene. Furthermore, a target sequence may have high homology elsewhere in the genome. These off-target...Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome Biol 19...Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 . https://doi....

...depositing lab. (For our more popular libraries, you might receive a similarly-sized aliquot from an amplification...ll want to infect your cells with the virus–the right amount of virus, that is. Ideally, each cell will...Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9...

... #MolluscMonday: #Crepidula atrasolea juvenile, light microscope with polarizing filter. Development is...an extreme number of nerve cells and an unusually high level of RNA recoding in the brain. RNA editing ...DNA in the eggs to get transgenic animals. They might be immortal. Manihot esculenta (Blake Meyers) Cassava...

...million dollar sales pitch. Given this large sum, one might expect that applicants would work for months to ...research? This is the central idea. It should be at a high level, broadly understandable to scientists outside...engaged by your presentation, then you’re on the right track. Furthermore, practice getting back to your...

...data inside the ELN is reproducible, and only the highly restricted number of people who can access the ...practical to move all this data in bulk into a lightweight tool like an ELN. It is nevertheless critically...

...interested audiences quickly and concisely – video might be part of the answer. Problems with written publications...TedTalks are very popular, though they focus mostly on highly innovative ideas rather than on the day-to-day ...that provides links to the videos, keeping the copyright and control over the videos with the researcher...

...recombination at a frequency three orders of magnitude higher than identical targeting vectors delivered through...Other investigators who had been working to develop high efficiency gene targeting approaches improved on...repair, it has proven more challenging to develop high efficiency CRISPR-based approaches for the generation...

.... This over-reliance on the perfect story for a “high impact” publication can lead to the publication ...reproducibility issues in the biological sciences. I’d highly recommend listening to the panel discussion to ...Scientific Figures into Dashboards and Share Your Insights Who gives a tweet? 9 facts about scientists on...

... LITE system, which uses light to regulate transcription, also includes light-regulated histone methyltransferases... CA (2017) CRISPR–Cas9 epigenome editing enables high-throughput screening for functional regulatory elements...

...may be a good choice for targeting neurons, with high transduction frequencies in the central nervous ...been reported (Muraine et al., 2020). AAV8 AAV8 has high transduction efficiency in multiple tissues, rapid...AAV9 AAV9 has relatively broad tissue tropism and high transduction efficiency, and is an excellent choice...