...different components, but all contain a FRET pair, a genetic component, and a small molecule component. Early...developed voltage sensor with a genetic component. The VSD components have historically come from a variety...have been engineered to have kinetics comparable to its one component counterpart and is significantly brighter...cellular localization component is typically handled by either the fluorescent component (i.e., when the dye...tools have two main elements: a voltage-sensing component and a fluorescent output. The GEVI family of gadgets... a hybrid approach and utilize both a genetic component as well as a small molecule which we will describe...reporter and the mechanism of their voltage sensing component, which separates them into two distinct categories...

... as Cascade (CRISPR-associated complex for antiviral defense) complexes, while Class 2 use single protein...effector complex after binding to a target. Class 1 is the most abundant of the two, comprising about 90%...tools using a Cascade complex is tricky, as you need to ensure every unit of the complex expresses at similar...have similar compositions and mechanisms, with the biggest differences being the components present in ...in the Cascade complex. Figure 2: Schematic of a Cascade complex (blue subunits) with a guide RNA...primary effector complex but are defined by the accessory Cas proteins in the Cascade complex. Despite a tenuous...the Cas effector may have the same name but be completely different! They are merely linked by a similar...

...are compatible with all your components. Next, consider the best form for those CRISPR components based...introducing those CRISPR components into your target cells. Delivery of these components into mammalian cell...most effectively achieved with different CRISPR components. Select the appropriate CRISPR tools for your...will you be editing? Do you want to deliver the components as DNA, or would mRNA or protein delivery be ...of the first steps is to identify what CRISPR components you will need to deliver. Minimally, one or more... the best method for introducing those CRISPR components into your target cells.  Mammalian CRISPR expression...best practices for efficient delivery of CRISPR components. If you are new to CRISPR or your model system...

...information about the cell’s complexity. A cell with a high degree of internal complexity such as extensive membranes... prepare sets of controls to aid with compensation. Compensation is the process of correcting spectral...need to analyze a specific subset of cells in a complex population. Have you spent hours in the biosafety...cell’s size, granularity, and target protein composition. It is used for a variety of applications such...and detecting specific populations of cells in a complex tissue or sample (1). When combined with cell sorters...instrument collects information about the cell’s size, complexity and fluorescence intensity.   What flow cytometers... cytometers detect Cell size and complexity, measured using visible light scatter The optical system has...

...co-coregulator) partners. The coregulator ‘core’ complex is composed of a tightly-bound invariant group of proteins...coregulators exist as multiprotein complexes, a member of a single coactivator complex can serve as a rate-limiting...seeking to compliment these consortium-generated datasets with a systematic effort to compile NR- and coregulator-related...physiological processes are controlled by deliberate and compulsory actions instigated by the central nervous system...factors found in metazoans. The superfamily is comprised of 48 human and 49 murine members. NRs permit ...metabolic, and immune response programs. NRs also encompass one of the most successful targets for drugs currently...interact directly with NRs) exist in steady-state complexes with secondary coregulator (or co-coregulator)...

...iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing of transgenes ...are prone to incomplete silencing of reprogramming transgenes, which leads to incomplete reprogramming... heterogeneity between clones. This can complicate comparisons made between samples, i.e. transgene expression...efficiency of reprogramming when compared to other methods, even when compared to integrating viral approaches...less time to complete. Morphology is a simple and fast to evaluate. iPSCs should form compact colonies that...and have a higher efficiency of generating iPSCs compared to other delivery methods. Retrovirus has a cloning...lentivirus has similar rates of reprogramming when compared to MMLV-derived retroviruses, it is not as well...

.... The team at the Victoria Makerspace is composed of computer scientists, engineers, and biologists working... elegans as a computer model. When I first heard about it, I got excited from a computer scientist perspective...defects above 25 degrees. Computer scientists and biologists work together to compare the predictions about...interests in biology and computing. In 2009, he founded a team at UVic to compete in iGem, an MIT-sponsored... really epitomizes what the makerspace is. The computer scientists can get involved in modelling, the ...Developing these skills relies upon biologists and computer scientists learning and communicating with one...coding the molecular biology of the worm into a computer model that recapitulates how molecular biology...

...of oxidation (dehydrogenation). MitoTimers are composed of a Timer protein fused to the mitochondrial ...upregulate an alternative pathway or induce other compensatory mechanisms. How can protein function be dissected...’s lab, acutely depleted proteins from their compartment of action via sequestration. The strategy relies...outer mitochondrial membrane. Sequestration is accomplished by knocking down your endogenous protein of ...May 1;126(Pt 9):2102-13. Assemble multiple component plasmids from building blocks via Golden GATEway...GATEway Cloning Frustrated by the complexity of assembling recombination of transgenesis constructs? A new...cloning kit simplifies the cloning process for complex DNA constructs, particular for those involving ...

...different compositions of the Cascade complex. Created with BioRender.com.   The Cascade complex Cascade... an important component of the CRISPR Cas3 system. Most Cascade complexes are comprised of a combination...systems work using a multi-Cas complex called Cascade (CRISPR-associated complex for antiviral defense). Cascade... Cascade complex from CRISPR type I-E. Created with BioRender.com. Thanks to this complex, developing... It can be difficult to ensure comparable expression of all components of Cascade for maximum efficiency...Cascade-Cas3 plasmids in Addgene's collection! A “complex” system Cas3 is part of the Class 1 CRISPR family...from different species and will have different compositions of Cascade. Although seemingly ubiquitous in...

... pre-mRNA splicing in a compound screen. Their reporter system is comprised of two plasmids: Luc (intronless... Vacuolar Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B ...insights into the make-up of multiprotein complexes like the EPC1 complex.. Going one step further, they also...about it! Split Cas9 system The Cas9 protein is composed of an n-terminal DNA recognition domain and a ...sequencing or other readout. Cellular memory is accomplished by the use of sequence-specific enzymes (e.g...calculated (the BRET ratio), which allows for compensation of varying fusion protein expression levels....more powerful LID, the authors recently used computational protein design, phage display and high throughput...