...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...

...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, ...

... 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 ...

...administered, which leads to many complications, including the inability of some institutions to know exactly...Furthermore, 7% did not enforce these policies. It is unclear how many postdocs are at each of these institutions...are afoot for the postdoc population, and it is unclear how this will all shake out. In the short term,...would help our resource. In the medium term, it is unclear what effects the new laws will have on the postdoc...

...that went beyond obtaining “omic” data and had to include clear plans to connect Genotype to Phenotype thus... is to ensure that, in general, these labs all include materials sharing in their dissemination plans ... diversity in the social bonds that they form including genetic mechanisms that control who a female wants...of basic evolution of fungi and eukaryotes and includes the fungus that is killing frogs all over 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...

...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...

...depend on target and sample type. Example steps include: permeabilization, antigen retrieval, and sectioning...available, and your experimental goals. Example steps include: counterstaining, mounting sample on slides, and...you optimize your protocol, always remember to include controls to show that your antibodies are binding...antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share some of their...

...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...

...AAVs to broadly spread throughout an organism, including difficult-to-target tissues like the central nervous...and so on, although we often use abbreviated nomenclature (AAV1, AAV2, etc.) to distinguish the capsids...AAV pseudotyping. It has broad tissue tropism, including liver, muscle, lung, CNS, kidney, and retinal ...serotypes used to target the central nervous system, including neurons, AAV5 is generally the best choice for...

...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...

...afflicted by pervasive off-target effects. This conclusion was based upon numerous phenotype discrepancies...importance of using orthogonal methods to confidently conclude the function of a gene of interest. Tools for ...discussed below). Examples of these constructs include pGAPi, which allows easy creation of long hpRNA...experiment are now available on Addgene. These include the construct that allows for easy insertion of...

...various sources that are difficult to trace. These include the laboratory personnel, the serum, the cell culture...cross-contamination and due to poor lab techniques. These include reusing pipettes for multiple cell lines rather...Contamination! Tissue culture media often have indicators included in the media that turns yellow from lowered pH... periods and affect the total cell viability. Conclusion To sum up, it is always important to prevent ...

...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...

... 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...

..., introduced the “Three Es of Immunoediting” nomenclature. 1. Eliminate - Latent tumors are recognized...on immune system Conventional cancer therapies include radiation therapy (owing to contributions from ...blocking. Other molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific... factors researchers are beginning to focus on include: Cell heterogeneity Tumors are composed of a wide...

... Figure 1: Labeled diagram of an antibody including Fc, Fab, heavy chain, light chain, constant region...antibodies for experiments in a variety of fields including neuroscience and immunology. They can be used ...analyzed using techniques such as flow cytometry. Conclusion Whew! Now you know that antibodies can be used...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ...

...for plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!   ...destination vectors. Destination vector options include humanized wt Cas9, dCas9 (transcriptional repressor... Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable ...filler sequences. You also have the option of including a previously-designed tyr gRNA, which causes hypopigmentation...