...beer. Ha! We had over 60 people join the fun - including a lot of future Addgenies as shown in the photo...employees. We now have a team of three (myself included) that plan big company events and fun things to...people you spend 40+ hours with each week. In conclusion - get out there any have some fun together! Laura...at all really! We've had plenty of events that included a simple corn hole set and a cooler. Chances are...

... flexibility is their broad range of targets, including proteins, peptides, amino acids, drugs, metal ...biosensors are used in a variety of applications, including detection of environmental pollutants, drug discovery...aptamers for use in many entirely new applications including aptamer based fluorescent biosensors. Aptabeacons...-based instead of protein-based fluorophores, including: Environments with higher temperatures, since...

...you start reading, there will be a temptation to include every piece of information that was ever published...figures/schematics or tables. And don’t forget to include citations so that people can go back and read the...studies that reported mutations in that protein (including how frequently a mutation was found and the size...doesn’t have to be revolutionary, but you could include a few comments on where you think the field is ...

...the CRISPR workflow. For a multiplex sample, ICE includes visual representations of all detected edit types...indel.   For a complete guide to ICE analysis, including detailed explanation and examples, read the paper...of the DNA-targeting region of the guide RNA, excluding the PAM sequence. PAM Sequence - The Protospacer...stack Genome Engineering Solutions, which also includes our CRISPR knockout guide design tool, guide validation...

...an interview next week and it includes a chalk talk. What should I include in my chalk talk?" So what exactly...proposed approach to resolve the knowledge gap? This includes the overview and the individual aims. Describe...Howard Hughes Medical Institute. His interests include the quality control of secretory proteins in the...

...completely understood, but some possible reasons include: Metabolic pressures - it takes cellular resources...when designing a full gene. A non-exhaustive list includes: Codon abundance relative to cognate tRNA abundance...insoluble, nonfunctional globs of protein known as inclusion bodies that will generally segregate with the ...assay to make sure your protein isn’t forming inclusion bodies and is folding properly. References 1...

...inside the body. So we screened gas vesicles gene clusters from a diverse set of gas vesicle-expressing species...right: colony whiteness on inducer plates, white inclusions in cells with GVs via phase contrast microscopy...this was the Anabaena flos-aquae gas vesicle gene cluster because of its small size and its ability to produce...a time-intensive endeavor. That is why we also included an analogous set of transient constitutively-expressed...

...domain - VP64.  Later generations of activators include the SunTag system which facilitates recruitment...Addgene’s website. Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB fusion, both targeted...targeted to the promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors...expression in different species and cell types including (but not limited to) bacteria, yeast, plants, ...

...an X,Y axis to generate a dot plot (Figure 1). Clustered events are called a population.                           ..., like a box, ellipse, or circle, drawn around clusters to define populations. This shape is called a ...diameter and volume of the cell, thus allowing the exclusion of doublets and debris. The side scatter roughly...show frequency of events, instead of relying on clustered dots. In a contour plot, each line contains the...

... hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. In molecular..., PhD, and Jonathan Gootenberg, PhD, focuses exclusively on this space — developing tools for cellular...measurable cell toxicity – using Cas7-11. This included tests done in cell lines known to be more sensitive... — the ability to now make any small changes, including base edits, insertions, and deletions. But CRISPR...

...made smaller. Previous attempts to “shrink” Cas9 include the use of St1Cas9 (~3.3 kb) from Streptococcus... approximately 1350 amino acids in size, which includes SpCas9, and shortern orthologs approximately 1000...in mouse liver.  Two other smaller Cas proteins include Cas12b and CasX. While there are no papers yet ...capabilities of full-length SpCas9. This set of plasmids includes AAV-Cas9C-VPR for targeted gene activation. Split...

...but some examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place...resolve these issues is with electrophysiology. Concluding thoughts and moving beyond the mouse While it...applicable to the rat [6]. Technical differences include the ferrule implants and patch cables. One huge... at the Rockefeller University.  His interests include the role of learning and memory in opioid addiction...

...episodic memory, a special type of memory which includes autobiographical events (see Jin & Maren, 2015...visualization of the morphology of individual neurons, including dendritic and axonal extensions and, most importantly...some engineering, RABV and HSV, but also others including pseudorabies virus type 1, vesicular stomatitis... Addgene's AAV guide)  Commonly used serotypes include AAV1 – 9, with most of these serotypes capable ...future developments in the field of AAV vectors. Conclusion An ideal comprehensive toolset for neuronal tracing...

...known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs...proteins that behave as monomers in mammalian cells include: mTurquoise2, mEGFP, mNeonGreen, mScarlet(-I), ...motif, which is needed for Golgi localization, is occluded and therefore this protein is mislocalized. The...that the fusion behaves well. But this is not conclusive evidence that the fusion operates in the same...

...Several factors contribute to the popularity of GFP including (i) fast and complete maturation to functional...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley et al... conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa = 3.1) and... or media. These sources of ‘autofluorescence’ include riboflavin in the medium or flavin containing proteins...

...works. Advances in Cre-lox recombination have included inducible Cre, such as the tamoxifen-responsive...mediate recombination.) The deletions are mutually exclusive, as either deletion removes one of the other loxP...immediately following the promoter. The STOP cassette includes a mutant YFP that does not fluoresce, but can ...More variants have been created by other labs, including R26R-Confetti described in Hugo J. Snippert, et...

...for CRISPR genotyping. The most common options include mismatch cleavage assays, such as Surveyor™, T7E1...information profile regarding putative off-target sites including both the edit rate and the repair product sequence...algorithms do not take into account other elements, including those relating to DNA structure (e.g. epigenetic...options exist for unbiased off-target detection, including Digenome-seq (Kim et al. 2015) for in vitro analysis...

... molecule at all, leading to inconclusive results, erroneous conclusions, and wasted time. In fact, many...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from...specifically to the expected protein. Common approaches include using RNAi to knock-down expression of your target...

...organizations in a variety of science policy roles including fellowships, volunteer positions, and part-time... with a large set of other societal pressures including economic, cultural, and political. Science policy...science blogs. Other avenues for skill development include leadership of extracurricular organizations, working...opportunities. NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab...

...organism to make the complicated transition from a clump of cells to a fully differentiated body. In contrast...monitor cell interactions in diverse tissues, including lymph nodes, brain, liver, gut and skin. Two-...able to help answer using two-photon microscopy include: How do T cells and/or Neutrophils travel to the...probes when using a standard Ti:Sapphire Laser include: Blue-green fluorescent proteins like EGFP Yellow-orange...