...Delivery The RNP delivery method consistently leads to high on-target editing and low off-target editing rates...SpCas9 The closer your edit is to the PAM site, the higher the efficiency of HDR repair. Check out this post...post to see if an engineered Cas9 variant might work for your experiment. PITChing MMEJ as an Alternative...

...scientists, MTAs are important—they guarantee certain rights and protections for depositor and requestor alike... must be executed. The Golden Stopwatch Award highlights TTOs that accelerate science by processing agreements...

...Keima accumulates in autolysosomes, this ratio is high. When untargeted, Keima can be used to measure bulk...autophagy-related microtubule-associated proteins 1A/1B light chain 3B, or LC3. LC3 is a commonly used marker ...oligomerization Seeing red: simple GFP photoconversion Light sheet fluorescence microscopy Resources on Addgene.org...

...forward. Here are a few out-of-the-box ideas that might jump start your learning. 1. Create a peer mentoring...what’s a scientist to do with no live audience in sight? You could do a talk virtually for your peer mentoring...university or use your current contacts to ask who they might suggest you speak with. See this post that’s all...

...their secondary structure, palindromic nature, and high GC content. This instability can cause partial loss...through ITRs due to their secondary structure and high GC content, but some companies now offer Sanger ...example, CAG is a strong promoter that drives very high, ubiquitous expression, while the human synapsin...

...Baradaran says.  Baradaran’s path to science began in high school. “I had a great chemistry teacher in 10th...medicines can have on human health. As a teenager in high school, I knew that this is what I wanted to do,...ClonTracer Barcoding Library, a library that “enables the high-resolution tracking of more than 1 million cancer...

...and it is under copyright, you will need to obtain written approval from the copyright owner and will ...treatment conditions like drugs (Chemogenetics) or light (Optogenetics) to activate receptors? If you’re ...

... host-pathogen interface, previously developed a high-throughput neutralization assay to study the link...Pfizer or Modern vaccine. This sera was used in the high-throughput SARS-CoV-2 pseudovirus neutralization... KL, Nitido AD, Murali MR, Alter G, Charles RC, Dighe A, Branda JA, Lennerz JK, Lingwood D, Schmidt AG...

...the ‘bells and whistles’ – including microscale light emitting diodes, thermal sensors, and microfluidic...channels. These features enable the fibers to emit light for optogenetic purposes, measure temperature, study...construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused to mCherry to the...

...makes them want to know more. This could be by outright stating the big-picture problem, by asking your...science storytelling. You can introduce your results right at the beginning of your communication, use conflict...storytelling options, and they all can be effective in the right circumstances.   However, in the midst of all these...

...pBR322, pACYC, and pUC were developed to provide higher copy number vectors that could be used in these...updated and added new content to our eBook. Some highlights of the 2nd edition eBook include: new chapters...

...determine genome-wide translational frequency. Although highly useful, this technique requires extensive sample... nucleus to bind cognate untranslated mRNAs. The high specificity and resolution of this system are achieved...SunTag Catch up on Your GFP Background Choose the Brightest Fluorescent Protein Resources on Addgene.org ...

...ask is what to do with them. In some cases, there might be a mutation that disrupts the function of YFG....drivers. It is much easier to mate flies to get the right cell type expression as opposed to making a new ... my next post, I’ll explain the relatively straightforward process of generating your own fly lines. In...

...viral samples. Read on to learn how aqueous samples might be affected by dry ice, and specifically how dry... removing the caps, incubating the tubes in an upright position for 30 seconds, and then recapping the...to prevent any pH change, as we still observed a slight change in pH (from 7.4 to 6.8). It’s possible that...

...as virus? “Like all things in science, getting a high titer Cas9 lentivirus prep takes a bit of time and...lentivirus, we are providing lentiviral preps containing highly requested gRNAs including those targeting EGFP,...library has retained gRNA representation, produce high titer lentivirus from the library, ensure gRNA representation...

...but thought we could learn a little bit more about high throughput DNA prepping by trying out NEB’s new ...generating a wide range of insert sizes and also produced high yields of quality libraries from small amounts of...summarize my experience in my first blog post! I highly recommend doing the same if you ever go to a workshop...

...Director of Biology, Lianna Swanson, suggested that I might be able to attend the 10 Years of iPSCs Symposium...or tertiary, or non-existant). On the very first night of the iPSC symposium, a panel of thought leaders...Resources on the Addgene Blog Learn about Efforts to Fight Rare disease with iPSCs Read About Addgene's Outreach...

...of mutant target (as low as 20 pg or 0.02%) in a high background of wild type targets. Comparatively, ...M. A Digital PCR-Based Method for Efficient and Highly Specific Screening of Genome Edited Cells. PLoS...PubMed Central PMCID: PMC4735761. 6. Fu, Y. et al. High-frequency off-target mutagenesis induced by CRISPR-Cas...

...this context, but also specifically study it. They might be researching enhancer elements that control the...research describe the production of plasmids that might be useful for your work. The links below direct ...techniques, the posts below may steer you in the right direction! CAPTURE-ing Chromatin Interactions: Using...

... Directors brings complementary experience and insight to the now 10 member board at Addgene. Let’s meet...Suffolk University Law School. Karen graduated with highest honors from the University of Michigan, Ann Arbor...Ernst & Young, LLP, auditing insurance, finance and high-tech industries. He has earned both an MBA in finance...

... synthesized SARS-CoV-2 expression plasmids in a high copy backbone with a T7 promoter driving multiple...GAL1 promoters. These vectors are intended for the high level expression and purification of SARS-CoV-2 ... strengths of intensive screening and the use of high throughput platforms, Addgene and Ginkgo are striking...

...followed by higher stringency PACE, resulted in the evolution of TadA variants with much higher base editing...simultaneously with 54% efficiency, about 7 times higher than ABE7.10. However, the increased editing speed...

...complicated? Well, while the techniques are straightforward, the systems – the cells growing and producing...And on top of that you need to worry about getting high enough expression of your protein of interest. If...by using Addgene’s Blue Flame award to identify highly requested GCE plasmids. But they’d also love scientist...

...variant that efficiently infects neurons, directs high-level transgene expression, and has retrograde functionality...genetic tool delivered by the AAV.   AAVrg requires high titers for efficient retrograde transport. For recombinase-dependent... at the injection site (Fischer, KB. 2019). The higher the titer, the more likely you are to see spurious...

...their former interns to fill entry-level positions right out of college. Prior to March 2020, internships... a nationwide database of  STEAM internships for high school, college, and graduate students. In order...interview, inspect your technology, background, and lighting for a smooth performance. Most importantly, make...

...narrow down search results by adding more terms.  Weighting We determined the relative importance of different...given resource, and designed the search engine to weight search results differently according to where a...matches of a name in the depositor field will rank higher than matches of the same name in the publication...

...). As shown on the representative plate to the right, colonies with an insert-containing plasmid have... plates at 4C for few hours after the initial overnight incubation increases pigment precipitation, enhances...colonies. Take care in making your plates: X-gal is light and temperature sensitive and needs to be added ...

...interested in the same things you are on a deep level. Right away you have something in common. Namely, the scientific... field will know about grants and projects that might help your work Tenure can depend on your national...some of my best studies were designed over late night poker games at Gordon Conferences). You don’t want...

...zygotes at 18 hr post fertilization. They observed higher targeting efficiency than in iPSCs, but with the...scientists proceed with caution and with proper oversight from regulatory and ethics committees comprised... Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes at...

... read an 11-page Cell paper - and sometimes, it might actually take me less time to read the book.  Intuitively...science communication, but the more you use, the higher cognitive load you are asking your audience to ...like the professor who once decided that to teach high school students PCR, they first had to understand...

...antibodies that produce detectable signals like light or color. In this post, we’ll go over some of the...chemiluminescent reaction. Chemiluminescence, which is light produced by a chemical reaction, can be detected...Whatever the conjugate, the molecules must somehow be tightly linked to your antibody. This chemical coupling...

...working with an endogenous locus. The most straightforward way to turn off expression permanently is to...- this section is for you! Many of the tools highlighted in this section can be used to turn expression...

...have less complementarity to the gRNA, they have a higher propensity to reform double helices and decrease...measurements to determine which portions of Cas9 might be important for nuclease specificity. They found...PMC4176945. 4. Kleinstiver, Benjamin P., et al. "High-fidelity CRISPR–Cas9 nucleases with no detectable...

..., TdTomato was seen only in cells with GFP, at a high activation frequency of 56-93%. In the converse ...Cre-DOG lacks the toxicity that can be seen with high levels of transcription activation domains. Both...constructing split variants should be relatively straightforward, but it is also possible to create split versions...

...chronic myeloid leukemia. Bhang says the work highlighted the importance of developing more potent ABL1...clonal dynamics in response to cancer therapy using high-complexity barcoding." Nature medicine 21.5 (2015...

...launched an online career coaching program that might make her coaching more affordable for you. There...transitioning a scientist from academia to industry, you might want to consult the Cheeky Scientist. Their motto...Successful Industry Professionals”. This larger shop might appeal to some although most scientist coaches can...

...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 ...

...undergraduate research questions. The idea is straightforward. Instructors first select a multi-gene pathway...O'Connor (left with Lab Director Doug Warner on the right) has taught courses in biochemistry, molecular cell...Addgene Videos to Learn about Basic Lab Techniques High School Students Explore Careers at Addgene Resources...

...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...