... and LV have been used successfully in preclinical and clinical settings for ocular conditions. However...used in clinical trials, a variety of engineered ones have shown great promise in preclinical studies ...that I got into the field of gene therapy, more specifically AAV-based retinal gene therapy. The year was...treat an inherited retinal degenerative disease called Retinitis Pigmentosa. I quickly became fascinated... Gelsinger in an adenovirus-based gene therapy clinical trial gone horribly wrong. It brought the whole...scientists, doctors and patients hope are many more clinical successes for gene (and cell) therapies. One may... over 25 years ago when completing her PhD and medical training in Ophthalmology. Gene therapy was in ...

...illumination on biochemical processes that are crucial in the world of biological research, medical therapy, ...components, typically a protein of interest. Fluorescent dyes and ligands vary in their chemical structure...both organic tissues and cell cultures, opening biological interiors to many inquisitive scientists. Seeing...the dye and the endogenous target, and thus no genetically encoded tag is required. These dyes can also ...be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently reacts...more flexibility due to the affinity tag they typically bind to. Stability of fluorescence: For applications...thorough removal of unconjugated dye molecules are critical for accurate and reliable imaging and assay results...

...cells and statistically analyze cell movements. By using cells labelled by fluorescent chemicals or by other... steady state or under physio-pathological challenge. However technical specificities have to be taken...complex metazoans, rapid cell division and large scale cell mobility are essential processes during embryonic...organisms, where tissues structures are more stable and local movements predominate (e.g. a basal progenitor cell...stage, only cells from the immune system show wide scale mobility with movement from the bone marrow and ... studying other systems in which there is large scale cellular movement throughout an organism. Studying...mobility than the static microscopy images of chemically coloured lymphoid organs did in the past. How...

...SR, Bird A (2017) Radically truncated MeCP2 rescues Rett syndrome-like neurological defects. Nature 550...awareness and published his observations in a German medical journal. Decades later and after years of painstaking...silenced by a Lox-Stop cassette, resulting in the neurological deficits observed in Rett Syndrome. Restoring... tamoxifen-inducible Cre not only halted the neurological damage but completely reversed it, regardless...this interaction to varying degrees determine the clinical severity of different MECP2 mutations. More recently...the MECP2 protein is necessary to rescue the neurological defects of Rett syndrome (Tillotson et al., ...amount of protein to each brain cell to restore physiological levels. Much research is now focused on therapeutic...

...Escherichia coli, like the construction of a so-called “genomically recoded organism” (3) but its portability...mutant library generation in several biotechnologically and clinically relevant bacterial species. pORTMAGE...Pathology have developed an alternative method, called STARR-seq (self-transcribing active regulatory ...new ligands for orphan GPCRs is of significant medical interest, however efforts to do so have been hindered...human cell line expression. In a twist on the classical TANGO assay, Kroeze et al. developed a new approach...Ras oncogenes have been difficult to target therapeutically due in no small part to the central role Ras... least three independent gRNAs, each chosen strategically to maximize the likelihood of creating a functional...

...generated from this research. At both a practical and ethical level, scientists arguably have a duty to...). Advances in genetic technologies and other biomedical innovations promise an improved understanding...best way to shepherd technologies through thorny ethical issues, ensure that everyone will have the possibility... fruits of scientific research, and prevent technological advances from exacerbating existing inequalities...based at the Department of Genetics of Harvard Medical School, has a mission of raising awareness and ...the benefits as well as the personal, social and ethical implications of genetics. In these conversations...sensitive topics and uncomfortable places, from sticky ethical dilemmas to dark episodes in the history of eugenics...

...for every application. Plus, the photophysical properties of chemical dyes provide many advantages over...depletion (STED) nanoscopy. Scale bar, 2 μm. Inset shows comparison with standard confocal microscopy. Image adapted... W. (2011). Chemical Tags for Labeling Proteins Inside Living Cells. Accounts of Chemical Research, 44...If you take a look in a typical lab freezer, you’ll probably find lots of different plasmids containing...substrate. If the substrate was linked to, say, a chemical dye or other functional ligand, your protein of...self-labeling tools: Tetracysteine: The first reported chemical labeling tag, from Roger Tsien’s lab in 1998 (...any non-cysteine amino acid) that reacts with biarsenical dyes such as FlAsH (green, based on fluorescein...

.... The overlap here is that it is unethical to use scientifically unproven technologies to edit a human...human clinical trials. This model also demonstrates that when we are at the point of scientifically rigorous...throughout the history of medical research, there have always been people who unethically experiment on humans...Cas9 genome editing is not far enough along, scientifically, to be doing in humans; We have not had a broad... recently published a report on the social and ethical issues of genome editing and human reproduction...research into both novel and previously established ethical issues raised around mitochondrial replacement ...respective mandates, to tackle a controversial and ethically fraught rapidly emerging technology. We could ...

...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 ...