...process allows you to physically magnify a sample so you don’t have to optically magnify quite so much...compatible with conventional antibodies, chemical dyes, and genetically encoded fluorescent proteins (Gambarotto... and the physical properties of the sample itself. For example, cultured cells are typically easier to...promises an easier, more accessible way to image biological features previously only visible via techniques...techniques also require specialized instruments and technical expertise, limiting their accessibility (Prakash... bigger? In ExM, and its various derivatives, biological samples are enmeshed in a gel of a highly absorbent...be expanded to ~4-10x linearly! This expansion physically separates molecules while maintaining their relative...

...the tumor, this results in frequent physiological and morphological changes. Tumors differ in their genotypic...their level of progression, anatomical location, characteristic (solid localized tumors or systemic tumors...favorably attaining the equilibrium stage. This can be called “stalemate equilibrium” where the cancer stops ...either due to age, infections, or other strenuous biological changes. In contrast to the acute immune responses...themselves from attack. It is a region of incessant chemical activity, and consists of other cell types, especially...attack the cancer cells are also suppressed by chemical signals such as IL-8, IFNγ, and colony stimulating...contributions from Madame Curie), chemotherapy, and surgical removal of the tumor. However, these conventional...

...coverslip creates mechanical tension, and the luminescence colocalizes with the focal adhesion marker paxillin... used the reporter to visualize biologically relevant forces at focal adhesions in living cells in several...Sensors for Bioluminescent Readout of Mechanical Forces in Biological Systems. ACS Sensors, 9(7), 3489–3495...disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag antibody collection grows... forms that limit its functionality. To make biophysical and structural studies more feasible, the lab...properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They were able... Cereblon construct for crystallographic and biophysical studies of protein degraders. Nat Commun., 15...

...of human (and most eukaryotic) proteins localize to chemically distinct subcellular environments, including...sequence intended to localize them to the secretory pathway, a dark pool that is mislocalized to the ER can ...in the eukaryotic secretory pathway."Biochemical and biophysical research communications 430.3 (2013):...While Vader was referring to the evil side of a mystical "Force," this quote is equally applicable to many...microscopy experiments with fluorescent proteins (FPs) localized to compartments other than the cytoplasm. That...resident endoplasmic reticulum (ER) protein instead localizes to the cytoplasm? Or more troubling, what if a...properly, FPs enable researchers to investigate the localization and dynamics of fluorescent protein fusions ...

...transmitted transsynaptically). This is because the recombinant forms of these viruses typically used in ... became foundational later on when we studied biological processes, like how our stem cells are housed... thus, are a major part of the answer to many biological questions about the brain. To address this, molecular...enter cells and deliver a variety of cargoes, genetically engineered neurotropic viruses emerged as powerful... (Nassi et al. 2015). The recombinant viruses typically used in neuroscience research (e.g., AAV) are ...spread is useful for neuronal tracing, it has two practical limitations. First, these high levels of viral...starter cell types, as well as to only spread monosynaptically from these cells. These modifications take...

..., an optogenetically controlled phospholipase D (PLD). Now they present a set of variants called SuperPLDs...membranes, indicated by the PA probe colocalizing with SuperPLDx30. Scale bar 10 µm. Images adapted from Tei...components of the cytoskeleton. They provide cells with physical structure, separate chromosomes during mitosis...Akhmanova Lab published a new optogenetic tool for localized microtubule disassembly, Opto-katanin (Meiring...Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule severing. A) Schematic of Opto-katanin...Katanin is of course named for the samurai sword called a katana. Opto-katanin consists of two separate...a stable cell line. This design allows highly localized, rapid and reversible microtubule severing and...

...holding a physical model of the protein in your hand – you will understand the power of a physical model.... researchers use physical models of protein structure? We have been creating physical models of proteins...work closely with local researchers to learn about their research while creating physical models of proteins... your hand? At 3D Molecular Designs, we create physical models of proteins using 3D printing technology...and I encourage you to hold one. Why do we need physical models of protein structure? Structural biologists...problem with the development of Byron’s Bender – a mechanical device that could be used to bend 1/8th inch ... example, when Tom Steitz was presented with a physical model of his large ribosomal subunit, he kept ...

...data The result was a fusion protein called CalFlux VNT. CalFlux VNT is composed of Venus fluorescent ...and NanoLuc® Luciferase. CalFlux VNT responds to changes in intracellular calcium and is useful for direct... requires long acquisition times that may be impractical or infeasible for certain experimental setups... phototoxicity, autofluorescence, and perturb biological phenomena. The technique of coupling FPs with...RLuc8 moieties in their Nano-Lanterns with NLuc and called their improved creations Enhanced Nano-Lanterns...luciferase Many intracellular sensors of events like calcium release have been made using fluorescent resonance...generally consist of a responsive protein (e.g. calcium binding protein) fused to two fluorescent proteins...

... techniques in which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are...placed in a protein site-specifically to alter its function, mimic a biologically relevant modification,...proteins, you’re probably quite interested in the canonical 20 amino acids. But in your quest to learn more...modification, or add in fluorescent or chemically reactive side chains, all of which can help determine structure...structure and function relationships or, in more practical applications, allow a protein to be more effectively...was developed into an E. coli system, with the noncanonical amino acid in the media and the tRNAs and synthetase...use. First, and most obviously, you need a non-canonical amino acid, which is usually added to the media...

...ll use calcium biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) ...content of this article. Biosensors (‘biological sensors’) are biological tools that monitor a process or detect...and the chicken myosin kinase calmodulin binding region RS20. When calcium is absent, the construct does...a reporter signal. Reporter signals may be electrochemical or light-based, with luminescent and fluorescent...like arabinose or hydrogen peroxide or ions like calcium and zinc. Some biosensors contain a single fluorescent...does not fluoresce; when present, calcium induces CaM/RS20 interaction, thereby changing the FP’s structural... biosensors from Robert Campbell’s lab display calcium-dependent increases in fluorescence of up to 11,000...

...editor-based clinical trial for SCD in 2022 and Editas Medicine starting a Cas12-based clinical trial for... no longer an accurate descriptor; progress in clinical therapy is more fitting, so it is time to update... approved or have made significant progress in clinical trials.     Figure 1: Types of delivery...some solid tumors are currently in early-phase clinical trials and have shown promising preliminary results...introduction of the cancer-fighting component, CAR. Clinical trials are underway to explore CRISPR’s capacity...for these diseases was sponsored by Vertex Pharmaceuticals and CRISPR Therapeutics. Data shared from these...-Cas construct targeting the diseased allele specifically. Early-stage trials showed a high success rate...

...Institute for Biological Studies) Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum...and iGECI calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators...biosensors for multiscale imaging. Nat Commun 7, 12405 (2016). Qian, Y. et al. A genetically encoded near-infrared...Shemetov, A.A. et al. A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol...photoreceptors incorporating biliverdin IXα (BV) autocatalytically. Fortunately for probe developers and users...NIR FPs and biosensors can be imaged at spatial scales from sub-cellular to whole-body in small animals...performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova ...

...Volume: < 0.05 ml Skill required: advanced Intrathecal Intrathecal (IT) injections are probably the most advanced...location requires a lot of skill and anatomical knowledge. Intrathecal injections are used for a wide array...experiments. Some of these techniques are what we call “beginner”, or injections that are easy to pick ...easier of the two, but it actually requires more technical skill to place the needle in the correct vein ...institutional oversight is required. Restraint: mechanical restraint or anesthesia Location: tail vein or...you might after getting a vaccine. Restraint: mechanical restraint Location: thigh Gauge: 25–27 Volume...xenograft models. Restraint: back scruffing or mechanical restraint Location: back or hind legs Gauge: ...

...per gene approach that has been the historical standard. Specifically, in our recent publications, we used...these “variants of unknown significance” (sometimes called “VUS”) function is unknown. How can we make a decision... if the majority of information on each cancer clinical sequencing report includes rare variants that ...characterization For many years, we’ve hoped that large scale laboratory-based functional experiments that classify...from testing this hypothesis: There was no large scale, broadly available collection of mutant cDNA clones... variants across a large number of genes as systematically as possible. In developing these assays, we...tumorigenesis assays, drug epistasis assays, and morphological profiling can be used as generalizable strategies...

...understanding of complex biological processes. Many tools exist to perturb biological function in a direct...However, recent work has called this prevalent assumption into question. Specifically, CRISPR-generated mutants...limitations of any technique in the context of the biological question. For example, while gene knockouts have...makes traditional knockouts unattractive from a practical perspective. Therefore, techniques that function...Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi is expressed as inverted...cannot be used to investigate essential genes. Historically, gene silencing methods were thought to be afflicted...genes of interest to trigger RNAi activation. A typical RNAi experiment can be divided into three fundamental...

...their localization and expression. Qiu saw that inhibition of the PV neurons resulted in mechanical allodynia...Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research technician in Kaspar Podgorski’s ... post-transfection, excited at 505 nm (right). Scalebar represents 100 µm. Image from Abhi Aggarwal. ...fourth year Ph.D. candidate at the University of California, Irvine where she studies opioid use disorder...will image these neurons using GCaMP7, measuring calcium signaling related fluorescence during morphine ... disorder,” says Hubbard. Figure 3: 20x confocal image of the periaqueductal gray (PAG), with red...Elizabeth Hubbard.   Haoyi Qiu: Pain response in mechanical allodynia Haoyi Qiu is a M.Sc. student at McGill...

...of the FRET pair. Since many biological processes occur within the typical FRET range, FRET efficiency ...nanometers of each other, or measure the tension in the helical structure of spider silk, or the activity of a ...Fluorescence Resonance Energy Transfer. Unlike the typical excitation and emission of an excited fluorophore...donor fluorophore to the acceptor fluorophore. The typical steps in FRET are: 1. Donor fluorophore excitation... between the fluorophores and serves as a small-scale ruler to measure distances that are too miniscule...Experiments using intermolecular FRET probes are typically used for studying protein-protein interactions...with an intermolecular probe is often either impractical (for biomolecules that are not proteins) or would...

...the undesirable side products generated during chemical synthesis of these molecules. To make large amounts...will get you started (Obst et al., 2017)! It is specifically curated for protein expression in K. phaffii... on using K. phaffii, then the MoClo YTK Kit specifically designed for S. cerevisiae is at your service...plant-specific parts you‘ll need, including sub-cellular localization signals, plant markers, and reporters (Engler... arena! The Mammalian Toolkit was engineered specifically for the types of expression and integration ... AAV delivery. While Fragmid developers were specifically testing CRISPR tools, it should be easy enough... MK system was used successfully for protein localization and transport studies in HEK cells, recombinant...

...full support of local law enforcement, and, although there was at least one person vocally protesting us...Stephanie Hays, a researcher at the University of California Berkeley. It’s been half a year since the march...Alina Chan is a postdoctoral researcher at Harvard Medical School working on human artificial chromosomes....rains, it was weather befitting of the ominous political climate in which American scientists find themselves...don't line up with the scientific consensus and calling for billions of dollars in cuts to important research...science, I loved the idea and decided to go. At my local March for Science in Albany, NY, many tables were...scientist at the University of Colorado, Anschutz Medical Campus.  He studies viruses including HIV and is...

...Spatially Confined Persistent Diradical Emitter. Journal of the American Chemical Society, 145(25), 13615–13622... Andrew York, Maria Ingaramo, and their team at Calico Life Sciences recently set out to do just that ... many organisms) are opaque, which makes most biomedical applications of these tools difficult or impossible...some previous attempts at magnetic control of biological systems (Wheeler et al., 2016; Duret et al., ... were very small or were possible only in non-biological conditions. So, they decided to search for a ...removed (Figure 1). At first, Andrew and Maria were skeptical. The team was careful to test the possibility ...record of developing new microscopy tools for biological applications across a wide range of topics. He...