...of unionization continues to be a polarizing one. High-profile universities remain outspoken in their opposition...FLSA changes, including finding money in already tight lab budgets and even simply being able to categorize...salary perspective, while this is a move in the right direction, the increase notably still does not meet...Workshop 2016 At past symposia, FoR worked to highlight some of the problems facing postdocs and graduate...

... vivo CRISPR screens. AAV vectors can be made at high titers equal to those achieved with lentivirus but...less involved setup when working with mice and slightly less red tape for approval processes. Additionally... means you can customize your CRISPR screen to a high degree. This is particularly helpful when investigating...Breakefield, X. O., Maguire, C. A., & György, B. (2019). High levels of AAV vector integration into CRISPR-induced...screening by AAV episome-sequencing (CrAAVe-seq) is a highly scalable cell type-specificin vivoscreening platform...

...quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous quality control...vectors, and helper plasmids (Chadeuf et al., 2005, Wright et al., 2011). In fact, the majority of hits come...samples from 0.1-20% and discovered that, with a high enough sequencing depth, we could detect contamination...with exquisite detail (Lecomte et al., 2015). This higher level of quality control is recommended but not...Acids 4:e260 . https://doi.org/10.1038/mtna.2015.32 Wright JF, Zelenaia O (2011) Vector Characterization Methods...

...Addgene we've set out to fill this gap by both highlighting that researchers do learn MANY transferable ...the lab and by offering advice on areas where you might need some help. Today in our transferable skills...you will have a much better chance of creating a high-performing team - which is every manager’s dream... a weekly basis! The silence and maybe even the slight awkwardness can spur people to speak up. Once aware...

...using a high-speed ultracentrifuge. During the spin, the virions will form a band due to their high density...nationwide where you can obtain excellent quality, high titer rAAV (including Addgene!), but you can also...terminal repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material...Zen, Z., Espinoza, Y., Bleu, T., Sommer, J. M., & Wright, J. F. (2004). Infectious titer assay for adeno-associated...

... backbone might be more prone to recombination, while a library with a large backbone might be more difficult...to ensure the highest possible transformation efficiency. Using Stbl4 or other ultra-high efficiency cells...specifically calculated based on the size of and weight of your library plasmids. As long as you achieve...

...chains and two small light chains that stick out to form the “arms” holding the highly variable regions that...membrane proteins that may unfold due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab...References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput Cloning System for Characterization of...resources on the Addgene blog Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification...

..., and so on and so forth. This also serves as a light technical control - an RQ value of 1.5 means you...quencher to quantify specific amplicons.  Sybr has high background noise, due to the large amount of dsDNA...DNA and the length of the amplicon and it has a higher rate of false positives (Colborn, 2008; Harridan...number of factors will determine which approach is right for your experiment. You’ll have to consider:  ...analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc Hum Genet. 2014;80:7.21.1-7.21...

...cells to grow and divide only at the right times and in the right places. Checks and balances: protecting...proteins naturally lack areas that are accessible, high-affinity binding targets for potential therapeutic...therapeutic agents. Nevertheless, a glimmer of light remains for this elusive target. Recent improvements in ...results and customize the small molecules for a higher affinity fit. Kevan Shokat’s group at the University...

...now, 2 decades later, is it finally back in the limelight. On December 19, 2017 the FDA approved Luxturna... brought all these key elements together at the right time and place. But it is the hard work and persistence...immunogenicity is critical when systemic delivery of high doses of vectors are needed. Additionally, AAVs ...ultra-rare ocular diseases are being benched due to the high cost of clinical development. As a result, Luk Vandenberghe...8. Trapani I and Auricchio A., 2018. Seeing the Light after 25 Years of Retinal Gene Therapy. Trends Mol...

...project she needs one of you to lead. At first you might think, “I’m not a leader,” because you don’t see...you. How is this new opportunity much different? Might you already have leadership skills that are just...and develop your nascent leadership skills. You might just find that you’re more of a leader than you ...External collaborators For further advice on highlighting your transferable skills in that big job interview...

...ancestry groups rather than self-reported race, it highlights the myriad issues researchers need to reflect...associated dependencies across ancestry groups (right).    Seeking an explanation  First, they looked...guides from being designed that target regions with high genomic variability.   Moving forward It took a ...Stephanie M. Fullerton et al. Getting genetic ancestry right for science and society." Science 376, (2022): 250...

...excluding Starbucks. Taking a nerdy perspective, you might denote your search as follows: (my_house + coffee...math illustrated. Intersect (left) and set minus (right). Enhancers are regions of DNA that demonstrate...enhancers, check out the Appendix. Conclusion: Shedding light on dark matter Precise knowledge of which stretches... courtesy of beyondthedish (left) and biobabel (right). People commonly think of DNA as a linear polymer...meters long. You twist and roll the string into a tight ball. Now the two ends of the string, instead of...

... may be just the method needed to jumpstart the fight against antibiotic resistance. Advantages of CRISPR... despite this resistance gene being encoded by a high-copy plasmid. In the case of the genomically encoded...distinct beta-lactam resistance genes, again with very high specificity. This double phage system both favors...bacteria. In mixed populations, their phages were highly specific for certain bacteria, just like the CRISPR...new strategy can help us win back ground in the fight against bacterial superbugs. References 1. Citorik...

... species, Zetsche et al. hypothesized that Cpf1 might represent a functional CRISPR nuclease that could... cleaves relatively far away from the PAM, NHEJ might retain the PAM site. Therefore, if HDR does not ...possibly mediate HDR. This “second chance” mechanism might improve the frequency of desired HDR edits, but ...editing efficiency for the Cpf1 orthologs was only slightly lower than that of the widely used SpCas9 and ...that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1...

...has drastically increased, it would have been a tight squeeze for all of us to fit in Joanne’s back yard...discovered my hidden talent in Ladder Ball... there might be a possibility it was just beginner’s luck, but... (only if it’s cool with your legal department) right in your breakroom. 5. Throw some physical activity...you have a very intricate itinerary, there is a higher likelihood that something will go awry. Don’t plan... My grandma used the old adage “many hands make light work” all the time. She was a wise lady for many...

...shielded from quenching by aqueous solvent. This tightly-packed structure explains the importance of the...permutations within each color also exist with only slight variations separating them.  Please note that many... Improves folding at 37C, reduces aggregation at high concentrations T203Y Red-shifts the wavelength...biosensors here. Optogenetics: Scientists can use light to detect, measure, and control molecular signals... blog, you should! He's got great articles and insight on the newest tools and techniques in fluorescent...

... This powerful negative selection system ensures high editing efficiency; the only non-edited cells to...The simpler system of pCRISPomyces-2 displays a higher editing efficiency, perhaps due to its condensed...Streptococcus thermophilus, Cas9sth1 as a robust and highly active enzyme for transcriptional silencing Mycobacterium...efficiency of knockdown and developed new, more tightly controlled, Tet responsive promoters to avoid leaky...dCas9 variant as well. Furthermore, they provide a high-fidelity (HF) variant of the BE3 that further improves.../10.1038/nbt.3011 Cobb RE, Wang Y, Zhao H (2014) High-Efficiency Multiplex Genome Editing of Streptomyces...

...technology has become very popular as a rapid and highly efficient way to move DNA sequences into multiple...two new recombination sites (attL-left- and attR-right-, Figure 1). Under certain conditions, the attL ...’ll walk through an example experiment where we might use Gateway cloning to generate our desired constructs...another, the inserted DNA fragment stays in frame. High efficiency The positive (antibiotic) and negative...M. (1992). A Genetic Switch: Phage (Lambda) and Higher Organisms (Cambridge, MA: Cell Press). Additional...

...three scenarios, but a short treatment regiment might wipe out bacteria that are completely susceptible...concentrations of a drug, but will be killed by a higher concentration, or sometimes more exposure time ... corresponds to the level of susceptibility you might expect if a certain antibiotic were chosen for treatment...survive for millennia. What we can do, however, is fight smarter, not harder. We can utilize tactics like...antimicrobials to learn how scientists are using CRISPR to fight bacteria. From a societal standpoint, we can all...