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...various structured surfaces. Plasmids expressing fluorescent proteins will be used to tag individual strains...within the biofilms will then be monitored using fluorescence microscopy and FACS. In an additional approach...this condition is crucial. Plasmids expressing fluorescent proteins and a new two-photon laser scanning ...plasmids containing promoter regions that activate fluorescent protein transcription in response to river contaminants...water samples at specific locations, measure fluorescent signals from the sensor, and send a signal including...

...separate DNA or transforming cells with green fluorescent protein, these activities are limited to those...Typically, if you want to make a protein – say green fluorescent protein – you first need the DNA sequence that... instructions it needs to produce that green fluorescent protein through transcription and translation...students can easily sense, whether that’s bright fluorescent proteins that they can see, enzymes that can ...Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent synthetic biology education kit." Science Advances...

... to our website through the use of alt text to describe images in online spaces. Alt text is used by screen...example. Remember, our goal is to use words to describe the image in a way that helps visually impaired... your alt text, use all of this information to describe the image helpfully. You and your readers are ...learned a lot from listening to others' alt text descriptions, and we’d love to hear yours as well.  More ...write alt text for any image you’re struggling to describe in the results. And many journals accept alt text...

...are now available for fluorescent reporting and genetically encoded fluorescent voltage indicators (GEVI...potential of a cell, the more differentiated and quiescent it is, and the more depolarized (positive) the...anion-conducting channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization...sensor and a mutated version of the pHLuorin fluorescent protein. This GEVI features increased brightness...

...agglomeration of fluorescent proteins into one particle within the cell, the lab fused the red fluorescent protein...(MAGIC). With MAGIC, scientists introduce an Escherichia coli donor strain into the microbiome to deliver...different bee species. BTK can be used to introduce fluorescent markers to visualize localization and distribution...division counting (DCDC), relies on the assembly of fluorescent proteins into one bright particle that segregates...

...genes across diverse cell lines.   dCas9-VP64 Description CRISPR activation can occur through fusing dCas9...activation. Synergistic Activation Mediator (SAM) Description SAM uses specially engineered sgRNAs to increase...VPR and SunTag) (Chavez et al., 2016). SunTag Description Rather than using a single copy of VP64 per each...expressed consistently throughout cells. VPR Description VPR fuses a tripartite complex with dCas9 to ... Signal Amplification in Gene Expression and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/...

...spreadsheet Data Field Description Plasmid Name Use a descriptive plasmid name and match the... in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options ...does this plasmid do?” in <200 characters. A descriptive purpose focuses on the practical application ... Primary Vector Type The vector type should describe how your plasmid is intended to be used. Choose...

...in this post: Pooled library to study antibody escape in SARS-CoV-2 Recombinant antibodies for aminoacyl...New ready-to-use viral preps Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz...SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom’s lab at the Fred...From Starr et al Cell., 2020. (B) Mapping antibody-escape mutations with the yeast display library. From ...characterized them by ELISA, homogeneous time resolved fluorescence, and suspension bead assay (Figure 2). After...

...co-express your gene along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow...incorporated into plasmids for research purposes are described below. IRES Elements Translation in eukaryotes...co-express your gene of interest along with a fluorescent protein or selectable marker, it is easiest to...genes at equivalent levels upstream of an IRES fluorescent reporter for easy detection.5  Just released:...efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. Kim...

... FAST system, Ye’s lab could edit a tdTomato fluorescent reporter gene in mice using Minicircle DNA vectors...of Cas9 and a photo-dissociable dimeric green fluorescent protein (pdDronpa) (Zhou et al., 2018). pdDronpa... flanking the DNA-binding cleft of Cas9, the fluorescent domains dimerize and prevent DNA binding in the...chromatin architecture (Hoffman et al., 2020). Fluorescent dCas9 proteins can be used to identify specific...gRNA and a dCas9 fusion protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9...figure below. Next, by using a dual reporter fluorescence assay, Hemphill et al. demonstrated that the...

...until new inflorescences begin to grow. This will also increase the number of inflorescences overall, ...and C terminally tagged fusion proteins with fluorescent (YFP, GFP, CFP, RFP, etc.) or affinity tags (...transformation, you can cut off the primary inflorescence (the flowering portion of the plant) near the.../books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September 2018). Additional resources...

...sample. A variety of chromogenic, chemifluorescent, and chemiluminescent substrates are available. Labs ...standard plate reader as opposed to chemifluorescent, and chemiluminescent methods that require specialized...peroxidase (HRP) or alkaline phosphatase (AP). Fluorescent tags may also be used, but for simplicity we ...

...Rogers, a postdoctoral researcher at the Friedrich Miescher Lab of the Max Planck Society. Zebrafish (Danio...tissues such as blood cells or neurons can be fluorescently labeled, allowing researchers to observe changes...guest blogger Katherine Rogers from the Friedrich Miescher Lab of the Max Planck Society. Katherine W. Rogers...researcher in Patrick Müller’s group at the FriedrichMiescher Laboratory of the Max Planck Society in Tübingen...about zebrafish behavioral studies Download the fluorescent proteins 101 eBook Read our plasmids 101 blog...

...this technological gap, a trio of recent papers describe the development of CRISPR activation (CRISPRa)...packaging capacity than AAV. Such an approach was described recently by the Day Lab, who developed neuron-...High-throughput screening strategies Although the tools described above make CRISPRa more accessible to neuroscientists...The pooled nature of this experiment requires fluorescence activated cell sorting (FACS) to isolate differentiated... in vitro and in vivo applications, the tools described above make CRISPRa in the brain relatively easy...

...successfully and expressing the protein. For example, fluorescence microscopy images resulting from our experiment...was not successful due to the absence of GFP fluorescence from the living, viable cells. This result presents...positive control, the internal control vector, was described earlier. Once you are sure your conditions are...produces the expected effect, which is green fluorescent cells (Figure 3).   Figure 3: Expression of Plasmid...come from multiple, independent sources. We'll describe these a bit more below, but the papers cited in...

...several secondary antibodies, increasing the fluorescence signal intensity. However, you need to be careful...sorting based on a single (lineage) marker, while fluorescence-activated cell sorting (FACS) enables you to...off by enzymes, turning the resulting molecule fluorescent and membrane-impermeant. With each cell division...(Figure 4a), yielding a stepwise decrease in fluorescence intensity in a (histogram) flow plot (Figure...division half of the dye molecules are lost and the fluorescence signal weakens. (b) The heterogenicity of proliferative...

...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool...context-specific effects of UAG suppression, as described (6). To determine the stoichiometry of phospho-incorporated...phospho:non-phospho species on PhosTag gels as described above. Reagents and protocols Sep-OTSλ plasmid...-Sung, et al. "Expanding the genetic code of Escherichia coli with phosphoserine." Science 333.6046 (2011...al. "A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation...

...of working with fruit flies is their ability to escape their habits and live in the lab itself. Many of...Light Interval System, or FLIS, incorporates a bioluminescence protein into the chitin that can be activated...everywhere find and re-contain any specimens that escape during routine lab activities.  Here, we demonstrate...plasmid-based system for locating and containing escaped flies. Nature 4:1-22. https://doi.org/4.01.2022...

...process of mouse model generation simpler, as described in an earlier post on generating CRISPR mouse ... make. For example, large insertions, like a fluorescent protein knock-in, suffer from low efficiency ...with your desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region ...most of the commonly used cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (...'d use ivTRT to create the ssDNA construct as described in Figure 2.  You can also use Addgene plasmids...

...delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral vector background Until ...lentivirus-derived vectors are capable of transducing certain quiescent or terminally differentiated cells such as macrophages...interest is expressed together with a marker (Fluorescent protein or selection marker) allowing for selection...process is to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...