We narrowed to 884 results for: ica

...colleagues have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al...Verkhusha’s lab has generated a new genetically encoded calcium indicator using small monomeric NIR fluorescent...Tuning the Sensitivity of Genetically Encoded Fluorescent Potassium Indicators through Structure-Guided...colleagues present an improved genetically encoded temperature indicator B-gTEMP designed to meet those... B-gTEMP itself is a significant improvement over previous temperature indicators, it also demonstrates...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...They generated variants optimized for different applications (v82 has higher affinity for glutamate but slower...

...or macromolecule. Biotin can be added enzymatically or chemically. Enzymatic biotinylation occurs naturally...molecular biology applications for biotin and avidin. A short list of some other common applications and links...downstream application. Biotin labelled oligonucleotides can be used in a multitude of applications including...post-translational modification of exceptional specificity.” Trends in Biochemical Sciences, 24.9 (1999b... M. “Biotin: The forgotten vitamin.” American Journal of Clinical Nutrition 75.2 (2002): 179–180. PubMed...dye-labeled molecules on surfaces.” Journal of the American Chemical Society 125.27 (2003): 8074–8075. PubMed PMID...this highly specific post-translational protein modification. Most organisms have less than five enzymes ...

...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...with an abundance of protocols and examples of applications from across disciplines now available. In this...experiment or evaluating ExM images in a new publication.         Why Expansion Microscopy?  Traditional...techniques also require specialized instruments and technical expertise, limiting their accessibility (Prakash...

...present.  Purification reagent contaminants The reagents used during nucleic acid purification can leave...thiocyanate (used in many plasmid purification kits), which can dramatically reduce the A260/230 ratio at ...Selecting Your Plasmid Purification Kit Resources on Addgene.org DNA Quantification Protocol Recovering... and are ready to move on to your downstream application — but wait! Do you know how much plasmid DNA ...(Figure 1B). The A260/230 ratio can be a good indicator of protein contamination, but a number of other...researchers traditionally use the A260/280 ratio as an indicator of protein contamination. However, this ratio ...suggest that the A260/280 ratio does not drop significantly until protein levels exceed 70% of the sample...

... 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... 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 ... time advances in human genetics led to the identification of several disease-causing genes, including...

...dCas9-fusion proteins has dramatically increased the potential applications of CRISPR/Cas9 in genome-engineering... tools for all-optical electrophysiology A major challenge in the field of all-optical electrophysiology... the perfect voltage indicator and optogenetic actuator pair. Ideally, indicators should possess fast ...sensitivity. This hierarchical screen led to the generation of two enhanced voltage indicators: QuasAr1 and...temporal control over CRISPR/Cas9 mediated genome modification and gene expression. The plasmids associated... system widely accessible for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol ...expression Brilliant technologies adopt basic biological concepts and employ them in creative and innovative...

... editing in Escherichia coli. Acta Biochimica et Biophysica Sinica. https://doi.org/10.1093/abbs/gmab036...far, we’ve used the MUCICAT system in two ways: In our initial paper, we used MUCICAT to optimize the gene...industrial strain. Not only is MUCICAT better, it is faster. Using MUCICAT only needs 2 newly constructed...engineered cells for green chemistry and biomedical applications.   Yiwen Zhang entered the Sheng Yang Lab...integration using CRISPR-associated transposases” or MUCICAT. Here, we’ll describe each of these CRISPR systems...providing users with a complete editing process. The MUCICAT system While the above systems are great for experiments...chromosomal integration (ex: genome editing), our MUCICAT system which is based on the CRISPR-associated ...

...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., ...detrimental as too little, resulting in MECP2 duplication syndrome or Rett Syndrome, respectively. One ...

...regulator. Other methods for physical shearing exist as well, such as sonication and needle shearing. Essentially... whole genome amplification (by a mechanism known as multiple displacement amplification) even in the ...proteins! Many thanks to our guest blogger Jessica Sacher! Jessica Sacher is a microbiology PhD student at...Their Viruses." Chemical Reviews (2016). PubMed PMID: 27319741. 3. Simpson, David J., Jessica C. Sacher, and...This post was contributed by guest blogger, Jessica Sacher, a microbiology PhD student at the University...bacteria) may be the most numerous and most diverse biological entities on our planet, but we still know collectively...adding methyl groups, glycosyl groups, and other modifications to their bases in order to protect their DNA...

...them. An easy way to do this is gel purification. In gel purification, you use a voltage difference across...backbone. Sample results indicative of successful and unsuccessful ligations are indicated below. A successful...grow overnight cultures for DNA purification. The simplest purification you can do is a miniprep, but if... maxiprep. In these purifications, you generally lyse the bacteria; add chemicals to precipitate out the...that can be easily maintained in a variety of biological systems. Read on for an in-depth breakdown of...digests. Of course there’s much more detail and verification required for the process to work well, so let...backbone. Because you lose some DNA during the gel purification step, it is important to digest plenty of starting...

... microscopy technique to incorporate optical sectioning. Optical sectioning refers to the ability to extract...Super-resolution microscopy uses a number of optical and chemical ‘tricks’ to turn subsets of fluorescent ...Richardson, Director of the Harvard Center for Biological Imaging and a Lecturer on Molecular and Cellular...are a sports photographer at the Super Bowl, a medical technologist taking an x-ray, or a biologist imaging...excitation volumes to enhance spatial contrast (i.e. optical sectioning (2) or super-resolution (3). Fluorescence...before reaching the detector. The pinhole is specifically placed to only allow light from the focal plane...of a sample. Therefore, when a large number of optical slices are acquired all fluorophores throughout...

... sport athletes, as well as the ethical and health policy implications arising from sport-related concussion... years of exclusively writing lab reports and empirical essays had also stirred some of my more creative...begun my journey towards a career in science communication, an established field of inquiry with which ...University of Otago’s Department of Science Communication the following year and, having thoroughly enjoyed...straight on to a master's degree in science communication. The first year of the course involved a whirlwind...world of industry. Along this road, science communication graduates are often well-equipped. Many institutions...truly benefit others. Herein, a good science communicator—with an understanding of how to appeal to a ...

...transmitted transsynaptically). This is because the recombinant forms of these viruses typically used in ...able to replicate, and for a virus to cross a synapse from one cell to another, it must replicate. Modified...tracing, it has two practical limitations. First, these high levels of viral replication are toxic. Second... well as to only spread monosynaptically from these cells. These modifications take advantage of the fact... 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...

...desired modification. Finally, the positive clones are sequenced to confirm the desired modification and ...polymerase activity (for amplification), 3’->5’ exonuclease activity (increases amplification fidelity), and no... genome modifications. Occasionally, multimerization of the PCR primers can cause duplication of the primer...the desired modification have been identified (and confirmed to be free of primer duplications), final validation...used to mutate the PAM sequence (an NGG sequence critical for Cas9 cleavage), thereby rendering the resulting...and the absense of additional modifications.   Site-directed mutagenesis of Plasmids. A) Generation..., which flank the targeted region. Upon PCR amplification, the template is removed by DpnI restriction...

...recombination. Yeast Replicating plasmids (YRp):  These vectors contain an Autonomously Replicating Sequence (ARS...These vectors replicate as though they are small independent chromosomes and are thus typically found as a...not necessarily promote replication.  Selection markers for yeast Historically, scientists have utilized...mammalian vectors, will catch you up on the core replication and resistance features of yeast vectors and ...proteins yet they still share many of the same technical advantages that come with working with prokaryotes...includes but is not limited to: rapid growth, ease of replica plating and mutant isolation, a well-defined genetic...mutation. Yeast expression plasmids used in the lab typically contain all the necessary components to allow ...

...are used across species: replication, selection, and expression. Replication It’s usually not enough to...chromosome has a single origin of replication (ori), where DNA replication begins, so a bacterial plasmid...Eukaryotes are a bit more complicated. Yeast vectors may contain an autonomously replicating sequence (ARS), a...fast-growing populations of cells, the vector needs to replicate alongside its host, or it will be lost from the...compatible ori that will be recognized by its host’s replication machinery. A shuttle vector designed for unrelated...experiment. Yeast shuttle vectors have these replication sequences for their yeast targets as well as ...episomal vectors is to include viral oris to drive amplification, which helps increase cellular uptake, but this...

...systems have been used as biological tools for a wide range of applications in molecular biology or in...activity of CcdB. Applications of ccdB in plasmid cloning Perhaps the most widely used application of CcdB is...selection markers. Restriction modification systems Because restriction modification systems consist of two enzymes...types of plasmid addiction systems and their applications. The discovery of toxin-antitoxin systems Toxin-antitoxin...CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA gyrase and...construct. Other toxin-antitoxin systems and their applications MazE/MazF Aside from CcdB/CcdA, there are many...counterselectable marker to create unmarked genetic modifications in the yeast Pichia pastoris. By combining both...

...a tail, and when gap junction-mediated electrical communication is briefly inhibited in the entire worm...fluorescent reporting and genetically encoded fluorescent voltage indicators (GEVI) are ideal tools to...thing that comes to mind. However, ion-based communication is not just for neurons. Non-neuronal cells ...It's just that for non-neural cells, ion-based communication happens at a slightly slower scale.  All living...living cells with a membrane possess an electrical state. This state, known as the resting potential (... (Vmem), can be measured as the imbalance of electrical charge between the inside and the outside of the...bioelectricity is revealing the important role of bioelectrical signaling mediated by ion-channels and pumps...

...are currently approved or have made significant progress in clinical trials.     Figure 1: Types...editor-based clinical trial for SCD in 2022 and Editas Medicine starting a Cas12-based clinical trial for...choice. In sum, CRISPR therapies have made significant clinical strides since we first wrote this blog post... no longer an accurate descriptor; progress in clinical therapy is more fitting, so it is time to update...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...

...Nature chemical biology. 2021 May;17(5):509-18. Inoue M. Genetically encoded calcium indicators to probe...Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins...transients in living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to... Imaging neuronal activity with genetically encoded calcium indicators. Cold Spring Harbor Protocols. ...Measuring calcium signaling using genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;...studies, you can start by asking ‘What is the biological phenomena I want to observe?’ This information...