We narrowed to 849 results for: Eras

...fluorescent channels on the microscope. With the proliferation of fluorescent proteins in recent years, there...the latest fluorescent proteins, I maintain an interactive graph and table of the best fluorescent proteins...TagRFP to give a brighter signal than mCherry. In general, filter sets designed for Rhodamine / Cy3 will ... of the improved folding variants of EGFP like mEmerald or Clover is probably best; mNeonGreen is an even...variants, presumably due to the lower growth temperature. This also suggests, however, that there is no...want the brightest signal, you may need to try several proteins in your system of interest. Finally, in...set of proteins the green and red proteins are generally the most detectable and so should be used to tag...

... Addgene is not dependent on grants. Aiming to operate under a similar model for our Viral Service, we...see the value to the scientific community in accelerating the Viral Service. With their support we are...helps keep our focus on assisting scientists and accelerating research and discovery, while also pushing us...take on this project? Addgene’s mission is to Accelerate research and discovery by improving access to...exposure in the community, we saw the opportunity to leverage our recognition, further fulfilling our mission... methods, performs extensive quality control, generates  educational materials, and provides viral vector...webpage). While the new team took the lead on operationalizing viral production and distribution, this project...

...tools so that researchers around the world can accelerate their work. After all, as we like to sayat Addgene... plasmid. LentiCas9-blast virus can be used to generate Cas9-expressing cell lines, which can then be ...system as opposed to a single-vector system) has several benefits. First, the low viral titers typically...lab. We provide protocols and useful tips for generating your own lentivirus. Secondly, you can use a ...expression is typically cytotoxic, which makes generation of high titer Cas9 lentiviruses challenging. ...rate variability when using lentiCas9-blast to generate Cas9-expressing cells. To ensure that this growth...confound your experimental data, we recommend generating a monoclonal Cas9-expression line prior to performing...

...20, 2021. By Will Arnold and Shreya Vedantam Accelerating research by improving the availability of materials...pandemic in which scientists race to find vaccines, therapeutics, and other treatments to stem the spread of ...plasmids to help academic and industrial labs accelerate research and to help find a faster solution to...SARS-CoV-2 materials, it will not scale well to the several thousand plasmids from Ginkgo. To balance the need...time consuming, costly, and laborious QC for all several thousand plasmids at once. Any samples that are... any scientists who requested the plasmid. By leveraging our core strengths of intensive screening and...balance by modifying certain processes in both core operations. As it has been in much of the world, we have...

...reduces the need to generate “single-use” transgenic lines. Instead, researchers can generate and maintain transgenic...each protein of interest and, for in vivo work, generate a dedicated transgenic line. Using zebrafish as...albeit slightly offset due to the nanobody-GFP interaction). The modularity of the system provides researchers...likely to afford the ability to deliver more therapeutics to the CNS, and with reduced systemic side effects... the liver.    Fig. 2: Summary of the generational evolution of the AAV capsids (left) and transduction...off target. Across all loci tested, error rates overall ranged from 0.3% to 10%, in comparison to the 62...

...bacteriophages as bacterial therapeutics. This idea was incorporated into Novophage Therapeutics, Inc. in March 2009...co-founder and current VP of Business Development & Operations of Sample6, for advice on how to take an idea...the application was changed, but Sample6 still leverages the core engineering and scientific concepts to...moved on to become VP of Business Development & Operations. Starting a biotech company When and how ...commit to the company, the internet, crowd funding, federal funding, foundations, and investors are examples...to Dr. Mike Koeris (Co-founder and formerly VP Operations of Sample6) for taking the time to speak with...

...lab in Harvard's Department of Stem Cell and Regenerative Biology is to understand the basis for cardiovascular...use their TALEN kit to quickly and efficiently generate human stem cells edited to carry mutant versions... In the Cell Stem Cell paper, for example, we generated hepatocytes to examine the effect of a gene on... metabolism. The TALEN kit is quite useful in generating the mutant cell lines that one needs to do disease.... That technology had been used for decades to generate knock-out mice, but for various reasons it is ...sequence a large number of individuals and find several mutations in the same gene or in multiple genes...Ding et al. "A TALEN genome-editing system for generating human stem cell-based disease models." Cell Stem...

...gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment with CRISPR ...expression. Location and sequence are important considerations for designing your gRNAs. For indels, it's ...off targets. For CRISPRa and CRISPRi, these considerations are of roughly equal importance (target should...worry less about sequence optimality because you generally have fewer sequences to choose from). Finally,...non-homologous end joining (NHEJ) often leads to the generation of indels and thus frameshifts that disrupt the... the introduction of a specific mutation, one generally relies on homology directed repair (HDR) to incorporate...user may need to optimize numerous parameters to generate the desired edit. The wrench: Gene activation ...

...exciting revelation. Science in any field can generally be applied to improve our lives and solve global...scientific process can help us understand how new therapies can be developed and applied. The current pandemic...Typically has a Master's or Bachelor's degree Generally a short-term position (1-2 years) Often a gap-...who opted to join labs in cancer biology or neurodegenerative disease because they had close relatives suffering...environment and with supportive mentors, being open and vulnerable about my struggles and what I don’t know yet ...also found that it is more common among first generation students, women, BIPOC, and other minoritized...like these can be highly beneficial. I’ve made several close friends and found incredible mentors through...

...dTomato) that promote interactions between the FPs interfere with native interactions, preventing proper ...the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity and...higher order aggregates. This property can be tolerated in some applications (e.g. labeling of cells or...homodimers or higher order oligomers can be analyzed by several techniques. Gel filtration and SDS-PAGE under non-denaturing...reflect the likelihood that fluorescent proteins interact or oligomerize in cells? This question is largely...fluorescent protein’s propensity to homodimerize. Several studies have used the OSER assay to demonstrate...

...address these questions, biologists often use next-generation sequencing. Sequencing the 16S rRNA reveals the..., 2019). They called their method Metagenomic Alteration of Gut microbiome by In situ Conjugation (MAGIC...vitro and in vivo. They identified cells from 19 genera across four phyla that have taken up the plasmid...vivo studies. However, they only identified seven genera in in vitro experiments, possibly because in vitro...one cell during cell division. To achieve the agglomeration of fluorescent proteins into one particle within...decreases over time and the number of elapsed generations can be determined. The lab used DCDC to measure...measure growth rate of E. coli for over 10 generations and showed that this method can be used for both in...

... research focuses on protein innovations for therapeutic and vaccine design. We all want tools that expand...of us still do), so we are used to the idea of general interfaces and the freedom to test out lots of ...of the coupling system include: Wide fusion-tolerance — Click Biology tags should be similarly useful...one finds in cells. High yield of reaction — generating product without a lot of left-over starting components...bonds. Reactions also can happen at both body temperature and on ice, when we have cracked open the cells...cross-reaction with SpyTag/SpyCatcher, allows for iterative expansion DogTag/DogCatcher SpyTag/SpyCatcher...systems can also assemble vaccines, target gene therapy vectors, or lock together smart biomaterials. There...

...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...churn out antibodies against the injected protein. Several weeks later, blood from the animal is drawn and...antigen binding, the variable regions alone can be generated as a fusion protein. An ScFv is made up of the...your samples to a standard curve. This curve is generated using a serial dilution of a known amount of protein...for other experiments to study protein-protein interactions. Flow cytometry Flow cytometry is used to detect...2014.   In vivo uses For both research and therapeutic purposes, antibodies can be used to affect cellular...functional grade antibodies. Some of the research and therapeutic uses overlap: antibodies can bind to and inhibit...

... Overview of the CRISPR classification hierarchy. The hierarchy flows from Class to type to subtype to...the array of CRISPR technologies. Learning the general principles of this classification system will help...classifications were placed into a fairly simple hierarchy: class to type to subtype to variant. Despite ...seemingly complicated steps to arrive at this hierarchy, the main aspects of each class and type are easy...Class 1, and are the most common type of CRISPR overall. Type I systems degrade large sections of DNA thanks...most common. Figure 4: Overview of the general CRISPR type VI mechanism. The Cas13 effector binds...diversity of rare CRISPR-Cas systems with deep terascale clustering. Science, 382(6673). https://doi.org...

...specific to the acceptor In order for FRET to occur several constraints must be met. The donor and acceptor...appropriately for energy transfer via dipole-dipole interaction. The efficiency of FRET can be measured for a...FRET range, FRET efficiency is used to infer an interaction between the fluorophores and serves as a small-scale...As the choice of fluorophore pairs is a key consideration for a FRET experiment, the possibilities can...of fluorescent proteins available. Fortunately, several excellent reviews have collated data from various... typically used for studying protein-protein interactions by measuring the change in FRET efficiency, ..., FRET experiments can probe protein-protein interactions, measure the concentration or activity of small...

...Performance dCas9-VP64 activation is generally thought of as the “first generation” CRISPR activator. While it ...initially published in 2013 (Bikard et al., 2013, Perez-Pinera et al., 2013). In the years that followed, innovative...Performance SunTag performs better than first generation activators while showing lower activation levels...to recruit transcription factors. Performance Generally, VPR is found to have significantly higher activation...method, dCas9 is fused to CBP, an histone acetyltransferase domain capable of rearranging chromatin structure...588 . https://doi.org/10.1038/nature14136 Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein...

...circuits with functional relevance following a set hierarchy. MoClo originally developed from Golden Gate Cloning...organization schema MoClo can be broken down into three hierarchical levels: individual DNA parts, transcriptional...For example: in a cell there are proteins which interact with other proteins and with DNA, and these complexes...are multigene systems whose protein products can interact with the DNA that encodes the protein itself or... or other genes within the circuit. These interactions yield a cellular phenotype, translational output...can support a larger cellular engineering goal, several of which are described below. Multigene circuits...promoter for a given peptide can easily be tested by generating Level 1 TUs with unique promoters, and the ideal...

...your study is aimed at developing human therapies. If a therapeutic is the end goal, it is best to work with...Targeting is also important in the development of therapies when expression must be restricted to specific... crossing the blood-brain barrier.  Another consideration when picking ligands for chemogenetic studies...ligands. However, they retain their intracellular interactions and still set off the same pathways, giving ... (Saloman, JL, et al., 2016).   DREADD ligands Several DREADDs, including hM3D(Gq), have been derived ...& Gold, M. S. (2016). Gi-DREADD Expression in Peripheral Nerves Produces Ligand-Dependent Analgesia, as...

... In this article we will discuss general viral safety considerations and best practices for handling lab-grown... basic biological research as well as clinical therapy. These biological tools are useful because they... separate plasmid (envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein... and the same holds true for types of viruses generated in the lab. The four most common types of viruses... lab and require additional safety measures.   General viral handling safety   Containment and personal...resistant to ethanol and isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some...

...Evan’s team generated their Myc antibodies. At first, anti-c-Myc [9E10] was just one of several new tools...over the years, several alternative antibodies against the Myc tag have been generated in an attempt to.... However, it was (and still is) infeasible to generate highly specific antibodies for every potential...would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different synthetic...epitope within that region. You’ll notice that “generate a reliable antibody for protein tagging approaches...