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... most commonly used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include: pHluorin...capacity. Many of these FPs are fluorescent when ectopically expressed in most organisms. Furthermore, fusing...fluorescence. Therefore, FPs are used to study many biological questions. The two most common uses are: 1) to...For LSS proteins, the Stokes shift is ≥ 100nm. Specifically, these proteins are excited by UV light or blue...were prone to oligomerize. This may affect the biological function of the FP-fusion protein. Therefore,.... al. (2010) “Fluorescent proteins and their applications in imaging living cells and tissues”  Physiol...

...excited to announce we now offer an export license application for rabies materials! Some of the materials in...international requestors. We also offer export license applications for VSV and SARS-COV-1 plasmids. Export control...will need to provide a brief description of the applicable research project when purchasing export-controlled... applied to cover processing costs. Once the application is approved, Addgene then ships the material ...offered VSV and SARS-COV-1 plasmids under this application process since May 2020. We find this process ...whenever possible, as we cannot expedite the application process.  More information about export control...

...translational applications (mechanistic studies and gene therapy treatments for neurological diseases). ...neuroscience research is the lack of a method for genetically manipulating neurons throughout the whole brain... creates a template that’s suitable for PCR amplification using pre-designed primers. By using this library...astrocytes. Why screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but...throughout the body and is often difficult to surgically access. Therefore an IV injection is an ideal...pictures (see figure 4). It also enables single-cell anatomical study and neuronal tracing, aka determining where...

... antibiotic resistance genes so that you can specifically isolate bacteria containing your constructs ...common antibiotic resistance genes and which applications are best (or worst) for each antibiotic. The...Ampicillin Ampicillin, often just called amp, is the historical king of the antibiotic resistance realm. Amp ...immediately after it has been heat-shocked or electrically zapped. As a result, a shorter recovery period...is expensive (relative to amp), and for some applications, the benefit it offers is marginal. Fortunately...antibiotic resistance ultimately comes down to your applications and needs. The more niche and specific your ... antimicrobial agents in MIC trays. Journal of Clinical Microbiology, 23(5), 959–961. https://doi.org/...

...that, they also help us keep track of the tubes’ physical location in the lab. These barcodes additionally...  freezers depending on the sample type. It is critical for us to know which plate, rack, and freezer ...missing so that we can make a new tube from a replicate sample. The transformation After the DNA has ...plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows us to make...help us track each sample, but we also have a dedicated and detail-oriented lab team that works carefully...performs an average of 140 transformations. The lab typically transforms newly deposited samples from Monday...Excel sheet.3. Be specific  - When assigning a physical location to your samples, include as much detail...

...Summer Science Communication Series. We’ll be covering the basics of science communication, tips and tricks...science with others, and introducing science communication as a research field. And there will even be ...be a few tips for scientific communication (manuscripts, talks, etc…!)  Deep Dives You probably are familiar... Dives” series comes in! We’ll take a common application or concept and go beyond the basics - diving ...you’re ready for the next steps on some common applications, join us as we get into the weeds on some of...

... gives you a single level of control without modification - either Cre is there or it’s not. Cre-mediated...37 °C). Smart molecular evolution led to the identification of FLPe which has a temperature optimum of ...asymmetric target site core but its 8 bp length that is critical for Cre and FLP function. Because recombination...Recombination between the two target sites with identical sequence leads to inversion of the flanked DNA...this plasmid mix results in sparse labeling of cortical neurons and their synapses.   How can you...control   Figure 2: Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry...(7): 657–62. PubMed PMID: 9661200.  3. Cardin, Jessica A. et al. 2009. “Driving Fast-Spiking Cells Induces...

...parenting strategies and make important ecological indicators since species live in water and on land....protocol community! Asclepias syriaca and Asclepias curassavica (Gerog Jander) Milkweed plants are known to many... as the only Monarch butterfly food. They have chemical pathways generating interesting drug-like compounds...Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky) This is not a new experimental...plants and are unusual enough to be interesting biologically. They have economic importance as harmful weeds... expensive. They are using a cool fluorescent indicator system to rapidly identify recombinants. Arthropods...

... this post I’d like to delve deeper and look specifically at the characteristics and capabilities an ELN...facilitate true reproducibility. The following are critical: (1) connectivity to other data sources used in...likely stored in institutional and lab servers specifically designed to hold large amounts of data. Although...group’s research – it makes no sense and is not practical to move all this data in bulk into a lightweight...lightweight tool like an ELN. It is nevertheless critically important to be able to conveniently reference specific...with a particular experiment or project thus significantly enhances the ability of others to subsequently... both accept deposit of data associated with publications into one of five general purpose open source...

...expression limiting regulatory elements or use a non-canonical code (Gustafsson et al., 2004), such as Hoogsten...folding and stability, change post-translational modification sites and even affect protein function (Mauro...recognize multiple synonymous codons) may actually be critical for determining the rate of translation. The ribosome...10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon optimization in human therapeutics...synonymous codon usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org... GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal of Neuroscience...synthetic? How Addgene;s dataset reveals trends in biological innovation Resources on Addgene.org Check out...

...described above showed successful target modification, indicating that AAV is a good delivery vehicle for...overall packaging capacity but necessitating purification and co-infection of two AAVs. Cas9 orthologs...post-injection. Livers from these mice were histologically normal and liver injury markers were not increased...the AAV-SaCas9-sgRNA constructs mediate genome modification, but they did so without a substantial immune...enough to deliver with AAV. This system fuses catalytically dead dCas13b to the catalytic domain of RNA ...finding smaller Cas9 orthologs works for some applications, it doesn’t eliminate the need to deliver larger...

...efficiency. This can be done through domestication, which is the modification of a DNA sequence to remove a ...This post was written by Alfonso Timoneda, with significant contributions from Robert Hurt and Mohamed Soufi... the synonymous mutation of a codon. You can domesticate the sites yourself using PCR, but gene synthesis...gene synthesis, you can request the sites to be domesticated through the process, and request your sequence...high-throughput, we recommend transforming into chemically competent cells in a 96-well PCR plate. Always...Modular Cloning Plasmids 101: Modular Cloning Applications and Kits Plasmids 101: Golden Gate Cloning ...

...very little data on which might work in your application. It can be quite difficult to decide on which...be searched and sorted by gene/target, name, application, pass/fail rating, sample species, antibody species... how the antibody performed in the submitted application, along with a brief description of the experiment...experiment. If that data was included in a publication, you'll find that information in the report as well... section will vary slightly depending on the application — flow cytometry, western blot, immunohistochemistry...data showing how the antibody worked in your application. The submission process is easy, with simple,...

...window onto communication between the muscle and brain, which may have therapeutic implications, say Hyo ...metabolites that are dynamically secreted by muscle during exercise to meet the brain’s physiological demands,” ...protects the brain from depression. Mice that are genetically modified to produce extra PGC-1a1 only in their..., the brain is protected from stress-induced biochemical changes that could otherwise lead to depression...

...speak for you (and you for them) Communication Being able to communicate effectively is part of what makes...excellent Technical knowledge and creativity Become a “thought leader,” develop a technical “niche,” develop...productive. Take all the opportunities you can to be communicating about science with others. If opportunities ... (be on a speaker committee) Follow industry publications like FierceBiotech to “learn the business” Attend...

...clones, it’s wise to quickly assess whether a significant number of cells have been edited. For many experiments...enzyme screening for resistivity to digestion, indicating targeted edits.   Interested in this method...sequencing confirmation. All knock-ins A method applicable to knock-ins both large and small is a software...consider using a version of Cas9 that has been specifically engineered for reduced off target editing, such...Tao Chen, Jos Jonkers, Bas van Steensel. Easy quantification of template-directed CRISPR-Cas9 editing. (...and variant-aware in silico off-target site identification for CRISPR genome editing. (2019) Bioinformatics...Resources at Addgene: Read our CRISPRGuide Get practical help from our CRISPR Protocols Find gRNA Design...

...). Advancing from this invention to KJ’s clinical application in under a decade is an astounding feat ...double-stranded breaks in the DNA backbone, making them significantly less prone to indel mutations that plagued CRISPR... safety of KJ’s experimental treatment, it was critical to reduce off-target editing as much as possible... ABE8e) would be tested for their base-editing efficacy in cellular models of KJ’s CPS1 mutation. Every...of identifying a successful editor. For this application, ABE8e turned out to be most effective. Targeting...editors with increased activity and therapeutic application. Nature Biotechnology, 38(7), 892–900. https:...

...which to test the validity of your biological model and the technical performance of your assay. But, in... a more-typical scale of tissue culture while still offering the potential for true biological insight...components, the model system and the assay are just as critical for success. Sometimes, you might have a few positive...facilitating more rapid follow-up. We hope the biological community will find these reagents to be useful...

...al. "Systematic identification of signaling pathways with potential to confer anticancer drug resistance... therapies,” said Kris Wood of Duke University Medical Center. “While we do not yet have a comprehensive... of 17 signaling pathways that are frequently implicated in cancer cell proliferation, survival, differentiation... make the library usable for pooled screens. Applications of the cancer pathways ORFs kit Wood and his... he said. “It could [be used to screen for] morphological features, migration, or survival in different...

...Thousands of mutations have been implicated in cancer. Yet, the majority of them remain uncharacterized...annotate genetic perturbations that may play significant roles in cancer pathogenesis. Their work culminated... of mutations they identified included gene amplifications and fusions, missense mutations, and indels...mutations were found to be “neutral,” having no significant effect on cell viability compared to a wild-type...Approximately ~22% of the mutations, however, significantly increased cell viability, and these “activators...aberration drivers of pancreatic cancer." Nature communications 7 (2016): 10500. PubMed PMID: 26806015. PubMed...