...an experimental system comes with fewer ethical and logistical challenges than does working with animals...grown to include local school districts, professional education associations, and local organizations such...Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for Applied Plant...University. Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two global stock...preference for using animal systems to demonstrate biological concepts - and you have yourself a plant publicity...publicity crisis. The Arabidopsis Biological Resource Center (ABRC) set out to remedy that with the establishment...conducting teacher workshops, or staffing a booth at local science festivals, ABRC gets the word out about ...

...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or ...Zhang’s lab describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM... and it consists of three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating...distinct effector domains that work together synergistically, modeled after natural transcription activation...GeCKO, a lentiviral CRISPR library enabling genome-scale, knockout screening). As the Zhang lab's Nature ... is robust, specific, and can facilitate genome-scale gain-of-function screening when combined with a ...demonstrated in a paper in Cell that they could typically achieve 90-99% knockdown with minimal off-target...

...specific and much faster than any genetic or pharmacological manipulations on cells, and has potential use... platform for visualizing RNA production and localization Article contributed by Tyler Ford  Listen to...set of tools that can be used to visualize RNA localization in mammalian cells. These tools consist of cobalamin-binding...platform at Addgene. Braselmann E, et al. Nature Chemical Biology. 2018 PubMed PMID: 30061719. New tools...expression vector (pRSETB). Fabritius A, et al. Cell Chemical Biology 2018. PubMed: PMID 30220597. ACE system...

...reporter for calcineurin activity in yeast Article contributed by Leah Schwiesow Calcineurin is a protein...intracellular calcium levels within the cell. When levels of Ca2+ are high in the cell, calcineurin dephosphorylates...attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics and can serve...identify new antifungals that target calcineurin. Current calcineurin gene reporters are based on the bacterial...screening, calcineurin dynamics measurements, or monitoring in vivo. To overcome these technical issues, ...genetic tool for labeling just a fraction of a genetically-defined subset of cells. By labeling fewer cells...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe...

...example image). Point Scanning Confocal Microscopy Point Scanning Confocal Microscopy was the first fluorescence... microscopy technique to incorporate optical sectioning. Optical sectioning refers to the ability to extract...detector. The pinhole is specifically placed to only allow light from the focal plane to reach the detector...excluded. Parallelized Confocal Microscopy (Spinning Disk) Parallelized Confocal Microscopy (Spinning Disk...drawbacks of widefield and confocal microscopy. First, widefield and confocal microscopy project excitation...multiple focal planes of a sample may also be useful for providing fast imaging along with optical sectioning...respective focal plane. In sufficiently thin samples, deconvolution can outperform optical sectioning ...

...another critical control in biological experiments. Replicates can be technical or biological and are ... inclusion of biological replicates such as multiple mice in each group and C, technical replicates or...recorded would be due to natural biological variation. This information is critical for correctly interpreting... accuracy and therefore the statistical power of an experiment. Technical replicates are repeated measurements...meaningful results.  Biological replicates are distinct samples that are treated identically, used to assess...binds specifically to its target through the antigen binding site but also binds non-specifically to other...binds specifically to its target through the antigen binding site but also binds non-specifically through...

...report physical titer, which is measured by qPCR.   Physical titer is typically calculated by two ...particle typically contains one viral genome. Other than a theoretical maximum value, physical titers don... you? Physical titer: the concentration of viral particles containing viral genomes. Physical titers ...facilities report the physical titer of a viral solution (Figure 2). Because physical titers are used for...that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type... AAV2 has been reported to have a near-perfect physical-to-infectious particle ratio of 1:1 (Zeltner et...for recombinant AAV2, the same study reported a physical-to-infectious particle ratio of 50:1 (Zeltner ...

... John H. McDonald. "A practical guide to evaluating colocalization in biological microscopy." American...subcellular localization of your protein of interest The first step in analyzing the localization of your ...organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside a cell with... proteins of interest and examine them using biochemical methods thus removing the context of the cell...living cell. Analyzing differences in protein localization and transport between healthy and diseased states...for example, validate that the fusion protein localizes properly by comparing the signal from the fusion...protein and see who it is hanging out with via colocalization - aka “are they close?” After tagging your ...

...and sensing electrical potential. Thankfully, there is a tool to measure these electrical events! Voltage...essential to their use as biological tools. Why Use Voltage Indicators? Electrical signals are integral to...Sensors This system consists of a genetically expressed enzyme, typically on the cell surface, which can ...the chemical factor, of a naturally occurring voltage indicator. To give an example, a genetically encoded... tag, cage, uncage, and trap the chemical fluor with the genetically encoded component of the system to...needs to be cell type specific as well as specifically localize to the membrane, otherwise they aren’t all... all that useful. The cellular localization component is typically handled by either the fluorescent component...

...interaction between a bisarsenical dye ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine...systems consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with... as intact tissue. Some chemical labeling strategies such as the bisarsenical dye (Figure 1a) and FAP ...Ronald T. Raines. "Bright ideas for chemical biology."ACS chemical biology 3.3 (2008): 142-155. PubMed... at the Janelia Research Campus, Howard Hughes Medical Institute. Chemistry is dead, long live chemistry...fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer...synthetic fluorophores. Add a dash of DNA with an electrical jolt and cells become perfectly capable of synthesizing...

...the amount of a given factor within a biological system. Basically, biosensors provide quantifiable readouts...ions, calcium is by far one of the most popular, with dozens of sensors developed for it. Calcium triggers...the presence of calcium. This includes FRET-based sensors such as Twitch. While calcium is king in the ...while neuroscientists would like to monitor calcium specifically in their favorite neurons. This specificity...The calcium is strong here….I can sense it….sound like a plot to an biology scifi movie? Or maybe it’...biosensor?! You can do more than just ‘sense’ biological compounds and reactions; you can quantify them...pretty simply – they are systems that measure biological phenomena by generating an output which is directly...

...colleagues have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al. 2022)... Verkhusha’s lab has generated a new genetically encoded calcium indicator using small monomeric NIR fluorescent... response kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G...has higher signal to noise ratio and is better localized for synaptic glutamate response. They generated...lower affinity) and with a variety of tags for localization (PDGFR, GPI, and SGZ). Read more from the authors...indicators with improved activation kinetics and localization for imaging synaptic transmission. bioRxiv 2022.02.13.480251...select variants based on performance metrics specifically for two-photon microscopy. The resulting JEDI...

...discovery–a review of theoretical aspects and recent applications." Current Chemical Genomics and Translational...al. "Designing logical codon reassignment–Expanding the chemistry in biology." Chemical Science 6.1 (2015...the American Chemical Society 123.48 (2001): 12123-12125. Journal of the American Chemical Society. 7. ...laboratory, then FRET experiments are a viable biophysical path to answers. What is FRET and what's it...Fluorescence resonance energy transfer (FRET) is a biophysical interaction between two fluorophores. When the...acceptor) if the two probes are close together (typically 30-100 Å, depending on the probes). In fact, FRET... FRET can be used to derive a precise physical distance between the donor/acceptor fluorophores (Stryer...

...Edward S., et al. "Millisecond-timescale, genetically targeted optical control of neural activity." Nature...one that can be achieved using pharmacological or electrophysiological methods (Deisseroth, 2011; Mei ..., et al. "Next-generation optical technologies for illuminating genetically targeted brain circuits." ...halobium differing in pigmentation." Biochemical and biophysical research communications 78.1 (1977): ...Over the past few years, however, remarkable technological advances have taken researchers to the threshold...led by neurobiologist Karl Deisseroth, combined optical and genetic methods to photo-stimulate mammalian... is reached and the neuron fires, sending the electrical signal down the axon (Lodish et al., 2000). Most...

...types of replicate controls: technical and biological. In general, technical replicates can be thought of...independent variable. Replicate controls: technical and biological Reliable results are reproducible from...NOT independent and are typically derived from one source. Conversely, biological replicates ARE independent... more in depth information. Technical replicates An example of a technical control is the transfection...independent variable. Biological replicates In any given set of experiments, the biological controls are more...the observed phenomenon in your experiment is specifically associated with the independent variable you...plasmids, and discuss why these control plasmids are critical for the correct design of the experiment.  The...

...essential aspect of biological research that has defined our understanding of biological processes and developments...Organism. News-Medical. Retrieved on September 18, 2020 from https://www.news-medical.net/life-sciences...evolving and demanding field, requiring a variety of biological and molecular tools. One such tool is the model...developments. Model organisms often feature genetic, physiological, or developmental characteristics that make ...are five more model organisms that have left a historical impact on the scientific community. (And, check... and biochemistry. Certain strains have been specifically developed to thrive in laboratory settings. ... used to study human cardiovascular disease, specifically stroke and hypertension (Iannaccone et al., ...

.... "A genetically encoded fluorescent probe in mammalian cells." Journal of the American Chemical Society...). GFP is the most popular, most widely used genetically encoded fluorescent probe. Several factors contribute...probe, it is the first choice when selecting a genetically encoded fluorescent tag. There are, however, ...of these strategies works best for a specific biological system under anaerobic conditions needs to be...of GFP may interfere with the activity or the localization of the fusion protein. Also, in cases where ...adding GFP may alter diffusion rates. Smaller genetically encoded probes are based on cofactor binding ...tags are peptides that are engineered to bind specifically and with high affinity to fluorophores. Examples...

...non-bench research career such as law, medical/technical writing, clinical, regulatory, product development,...Further, this post may contain technical inaccuracies or typographical errors.  No action should be taken...industry H-1B There is a numerical quota of 85,000 total H-1B visas approved each fiscal year: 65,000 visas ...a "specialty occupation" as requiring theoretical and practical application of a body of highly specialized...a US Master’s degree or higher degree. These numerically limited visas are referred to as “cap” subject...chemistry, architecture, engineering, mathematics, physical sciences, social sciences, medicine and health...industry such as technology transfer, communications, medical writing, compliance, core facility management, ...

...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since...transfer (FRET)-based calcium biosensors. Perhaps most notably, the Twitch calcium sensors, which are available...-affinity for calcium binding and developed a version of the protein with as few calcium-binding sites...engineered sensors were designed to replace synthetic calcium dyes, which had been in use since the 1980s. “Synthetic...of interest,” Griesbeck said. Because they are chemical compounds, they have to be applied or injected...they shift from "off" to "on" in the presence of calcium. This color change can be observed under a microscope...Griesbeck and his colleagues began by isolating a calcium-binding protein, troponin C (TnC), from muscle ...

...signaling events occur in a physiological setting. Genetic techniques typically rely on overexpression or...needed for function. The lab validated this vector, called pBAM1, by conducting random mutagenesis in the ... sgRNA delivery and providing a means for large scale functional screens. For more information on these... post,Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening. Also, check out our updated...brighter mutant S65T-GFP, and the subsequent biochemical characterization, has enabled scientists to engineer...observations: Rapamycin is a membrane-permeable pharmacological agent that can rapidly induction heterodimerization...flexible cloning process provides improvements to classical cloning methods, particularly for complex transgenesis...