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...FRET Fluorescent Proteins: FRET FRET Pairs FRET Calibration Standards Additional Resources Förster (or Fluorescence...between the fluorophores at the single-nanometer scale, scientists can use FRET to report on protein binding... nm 4 ). Values obtained from FPbase.org FRET calculator (Link opens in a new window) *: Long Stokes shift...new window) for more detailed discussion. FRET Calibration Standards The articles or collections listed ... with positive and negative FRET controls or calibration standards consisting of donor and acceptor proteins...multiple FRET acceptors mTurquoise2-mVenus FRET calibration standards and fusion proteins Feldmann et al....new window) External Resources FPbase.org FRET calculator (Link opens in a new window) Nikon MicroscopyU...

...) domains that respond specifically to small molecules termed Pharmacologically Selective Effector Molecules...Plasmid Collection Chemogenetics plasmids encode genetically engineered receptors that are activated by specific... to artificially activate or inhibit specific biological processes. Addgene's collection contains two ...Activated by Designer Drugs (DREADDs) and Pharmacologically Selective Actuator Modules (PSAMs). DREADDs...have been engineered to respond to specific physiologically inert small molecules. There are four types...

... site into a genomic attP site. So far, their practical application has been limited by low integration...sequential strand exchange at a specific target site (called loxP for Cre, or FRT for Flp). The nature of the...other recombinase systems. Enzyme Target Site Canonical Sequence Structure (Length) Common Variants Cre...repeats + 4 bp spacer (32 bp) Currently, only the canonical rox site is widely used. Flp FRT GAAGTTCCTATTCCGAAGTTCCTATTCtctagaaaGTATAGGAACTTC...Expression: A stop codon flanked with loxP sites (often called a "lox-STOP-lox" or "LSL" cassette) placed upstream...Placing the loxP sites on either side of a gene (called "floxing", for "flanked by loxP"), will permit ...excise the shRNA to return gene expression to physiological levels. In lox-STOP-lox shRNA constructs, Cre...

... of stochastic combinations of cell-filling antigenically distinct protein tags (or "bits"), creating ...well as barcoding with specific enhancers for morphological readout. How to Use PRISM Barcodes are delivered...AAVs either directly to a region of interest or systemically using a capsid capable of crossing the blood-brain...microscopy workflows to achieve high-resolution optical imaging down to individual synapses. Figure 2: ...new window) , and The Max Planck Institute for Biological Intelligence (Link opens in a new window) . Read...barcodes enable self-correcting neuron tracing with nanoscale molecular context . bioRxiv 2025.09.26.678648;...

..., excitatory and inhibitory opsins , genetically-encoded calcium indicators, and rabies targeting genes...doubly-specified combination of genetic and/or anatomical-defined parameters, by placing two orthogonal...fragments (which have become exons) determines the logical expression requirements of Cre and Flp (B,E). When...Fenno LE, Ramakrishnan C, Kim YS, Deisseroth K, Callaway EM, Oberhuber M, Conzelmann KK, Staiger JF. 2019...Parvalbumin-Expressing GABAergic Neurons in Barrel Cortex Reveals Local and Long-Range Circuit Motifs. Cell Rep. 28(13):...Robertson K, Osten P, Sabatini BL. 2019. Distinct Cortical-Thalamic-Striatal Circuits through the Parafascicular...

... use a mechanical method before your enzyme lysis. For cell cultures, enzyme lysis is typically sufficient...for your experiment. But if you understand the technical aspects of a western blot, you'll be able to understand... so let's dive in!      Figure 1: The technical decisions to make when designing a western blot...multiplexing is challenging and limited. Direct westerns typically rely on primary antibodies conjugated to fluorescent... antibody or loading sample, which may not be practical. Conjugated primary antibodies can be more difficult...lengthy protocol, require the use of hazardous chemicals, and are not always consistent. Precast gels, ...unless you buy specialty nitrocellulose membranes specifically developed to allow stripping. And as a safety...

...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously...reassemble when exposed to specific UV wavelengths or chemicals (Polstein and Gersbach, 2015; Zetsche et al., ...much cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based research...pathways and genetic interactions. Additionally, large scale CRISPR-based screens can be used to gain insight... Tuttle and Alex Chavez! Marcelle Tuttle is a medical student at Tufts University and former researcher...researcher at the Wyss Institute for Biologically Inspired Engineering.     Alejandro (Alex) Chavez is a member...MC, Qi LS, Kampmann M, Weissman JS (2014) Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation...

...’s disease (PD) is a chronic and progressive neurological disorder that primarily affects movement. More...-wide association studies (GWAS) or other large scale studies. Researchers are now investigating the role...3xFlag-eGFP-Flag-SETX construct for subcellular localization studies with GFP, or IP with Flag tag. (Bennett...organelles. (Mattia et al. EMBO Rep. 2018) Study intrinsically disordered protein regions with FUS and HNRNPA1...fluorescent proteins, including Markers of Subcellular Localization , Optogenetics Plasmids , and Chemogenetics ...(Link opens in a new window) A collection of neurological mouse models including complex and monogenic...

...with most experiments, you first need to have a biological question in mind. Then you can decide how to ...additional sites throughout the genome. These sites are called off-targets. Be aware of off-target sites when ...protein and gRNA, which you generally perform by chemical transfection or electroporation. Expression can...questions: Does the resulting population need to be identical (clonal) or is a mixed population okay? Does the...from a single cell and will result in cells with identical genomes. If your end goal is to establish an edited...Perturb-Seq: Dissecting Molecular Circuits with Scalable Single-Cell RNA Profiling of Pooled Genetic Screens... Kampmann, M., & Weissman, J. S. (2014). Genome-Scale CRISPR-Mediated control of gene repression and activation...

...newly developed calcium sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs...biosensors. Nature chemical biology. 2021 May;17(5):509-18. Inoue M. Genetically encoded calcium indicators to...Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...fluorescent proteins linked to calcium binding proteins (e.g., calmodulin or troponin C). Their fluorescent... monitor calcium concentrations and transients in living cells and organisms. Unlike chemical indicators...unusually fast calcium dynamics, whereas non-neuronal systems tend to have slower, but larger, calcium changes...sensitivity parameters. Sensitivity and Localization The calcium concentrations in different subcellular...

...values – numerical risk allowance in a statistical test. Distribution - spread of data; typically, normal...Statistics – The basics Statistical analysis is what determines if there is statistical significance among ...are statistically different. They are classic parametric tests and one of the simplest statistical tests...if the dependent variables worked synergistically, antagonistically, were additive, etc.). ANCOVA and...analysis can provide information on specifically which groups are statistically different within your ANOVA/ANCOVA... data in ascending numerical value relative to quantiles derived from a theoretical distribution. If your... non-parametric statistical tests! Adding it all up  There are many, many statistical tests beyond the...

...names for technical support roles. Examples are Technical Service Representative, Technical Support Specialist...these departments from technical support. Technical support careers in brief Technical support careers are...troubleshoot problems at the bench. Specifically, roles in technical support make excellent use of the ...worked as a technical support specialist for 4 years. Read on for insights from my time in technical support...products successfully. In technical support you may have to answer technical enquiries such as: “Can I... areas of expertise in technical support positions. When applying for technical support positions, you...familiarize yourself with technical support. Even having personal experience with technical support through the...

..., which allows you to visualize the cellular localization of your protein. Additionally, tags are...expression Fluorescent proteins (GFP, mCherry, etc) Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression...the Michael Davidson FP Collection NLS Nuclear localization dCas9 plasmid - dCas9 expression plasmid without...pEntry vector that adds a C-terminal nuclear localization signal pGADCg - Gateway yeast expression vector...vector that adds a SV40 NLS Myr Membrane localization pWZL-Neo-Myr-Flag-DEST - pDest vector... the plasmid. Selectable Marker Typical Host Organism Representative Empty ...

...monitoring endogenous protein dynamics in highly scalable assays. HaloTag Fusions HaloTag (Link opens in...HaloTags form a covalent bond with these synthetic chemical ligands, which have been developed for special...levels in lysates or live-cells, thus preserving physiological relevance. NanoBRET Assays NanoBRET (Link opens...similar premise, but examine the proximity of a chemical compound and its target protein. The apparent ...Connections (Link opens in a new window) blogs for technical tips and tools for genetic reporter assays (Link...

...choice of seven serotypes and four volumes. Details Scale and Titer Packaged on Request can be ordered in ...preparations, we set both minimum standards and typical titers, though actual yields may vary by construct...construct. Size Quantity Total Volume Minimum Titer Typical Titer Small NEW 2 × 100 µL aliquots 0.2 mL 4 × ... with a minimum titer of 2 × 10 12 GC/mL and a typical titer of 7 × 10 12 GC/mL. For more details, see...providing scientist through an MTA. This process typically takes 1–4 days, depending on your institution....

...following regions: Isocortex Hippocampal formation Cortical subplate Striatum Pallidum Hypothalamus Cerebellum...pAAV-AiE2051m-minBG-SYFP2-WPRE3-BGHpA AiP1277 AiE2051m SYFP2 Claustrum Cortical subplate 220711 pAAV-AiE2051m-minBG-iCre(R297T...R297T)-BGHpA AiP1786 AiE2051m Cre Claustrum Cortical subplate 230518 pAAV-AiE2051m-minBG-FlpO-WPRE-HGHpA ...pAAV-AiE2051m-minBG-FlpO-WPRE-HGHpA AiP1211 AiE2051m FlpO Claustrum Cortical subplate 191720 pAAV-AiE0743m_3xC2-minBG-SYFP2-WPRE3-BGHpA ...and transgenic mouse lines for genetic access to cortical cell types. Cell , 188 (11), 3045–3064. https:...for Brain Science BRAIN Armamentarium Collection Caltech Systemic Capsids AAV Resources AAV Packaged on ...

...horizontal stripes on the sides of the adult body, is a tropical freshwater fish in the minnow family. It has long...inflammation and infection, regeneration, human neurological disorders, microbiota studies, and drug discovery...Cell lineage tracing in multicellular systems typically relies on genetic markers, somatic mutations, ...types. CreLite - George Eisenhoffer Lab. An optogenetically controlled Cre/loxP system that enables precise...assessment of lipoprotein abundance, size, and localization in high-throughput. MAZERATI platform - Leonard...

...expression, protein localization, protein–protein interactions, and other physiological processes in plants...repository contains many plasmid tools that were specifically designed for use in plants. These resources ...Infrastructure Kit Modules connect MoClo and Gateway for hierarchical assembly of plant multigene constructs; contains...an Old Favorite External Resources Arabidopsis Biological Resource Center (ABRC) (Link opens in a new window...

...from University of Calgary. Over the past few decades, genetically encoded calcium indicators (GECIs) ...response to calcium, which is more than just the stuff that makes bones strong; it plays critical roles in...}   Abhi Aggarwal is a medical student at the University of Calgary and a research associate with...Vectors 101: Calcium Sensors New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor A Practical Approach to...and cell signaling throughout the body. Because calcium levels change rapidly during neuronal activity,...our toolbox, our team recently developed a new calcium indicator, OCaMP. The challenge: Expanding the ...from FPbase.org. Scientists have developed calcium indicators spanning nearly the entire visible spectrum...

... the types of viruses used as biological tools.   Viruses as biological tools In nature, viruses infect... why viruses are such wonderful scientific and medical tools. As scientists, we can manipulate what this...effectively infect a wide range of cell types that are typically ‘difficult to transfect’, such as primary cells...with whole organisms. Finally, viral delivery is typically straightforward across applications, while electroporation... cells. They accomplish this through nuclear localization signals which can traffic their viral particles...Packaging Capacity (~4.5kb From basic science to clinical applications, viruses are true bench to bedside...