We narrowed to 695 results for: prot

...probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering what the...RNAs (ncRNAs) are transcribed but do not encode proteins. They play important roles in many cellular and...functionally manipulate and study in comparison to protein-coding genes (Rinn and Chang 2012). Scientists...Scientists have determined the function of many protein-coding genes thanks to systems like CRISPR/Cas9 that allow...often used to knock-out, or permanently disable, protein-coding genes by generating insertions or deletions...powerful tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription...lncRNAs and has fewer off-target effects. Newer Cas protein variants may also be useful for targeting ncRNAs...

...indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (e.g., calmodulin or troponin...Many calcium reporters use a single fluorescent protein (FP), requiring only a simple measurement. However... mechanism-guided design of single fluorescent protein-based biosensors. Nature chemical biology. 2021...encoded calcium indicators. Cold Spring Harbor Protocols. 2012 Jun 1;2012(6):pdb-top069609. Palmer AE, ...genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources on Addgene.org....org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’s Viral Services  Resources on ...

...tags to the C-termini of Met proteins, which can be used to confirm protein expression by western blotting...function wide open. What if two seemingly similar proteins have acquired divergent functions due to mutations...their experiments, students test whether or not proteins from the fission yeast, Schizosaccharomyces pombe...deposited with Addgene, to successfully analyze Met protein complementation. In the procedure diagrammed in...manuscript in preparation), indicating that Met protein functions have been largely conserved since the...Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...TadA7.10 in ABE7.10)  a catalytically impaired Cas protein (dCas or Cas nickase)  a guide RNA that targets...to ABE7.10. Depending on the position in the protospacer, the editing capacity of the ABE8s ranged from...more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase the targeting scope of adenine base...site in the genomic DNA to contain a specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N... Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG) to create NG-ABE8... genomic editing by measuring the reduction in protein expression of these six genes. All ABE8s edited...efficiently than ABE7.10 with ABE8.20-m reducing protein expression the most in primary human T cells. While...

...figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning method procedure...gene of interest (or, if your protein is fused with a fluorescent protein), fluorescence microscopy can...on Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression... place to screen your clones. Depending on the protein, we have used Western blots, dot blots, ELISAs,...

...fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent protein tools. Future...to creating and sharing many great fluorescent protein tools, Professor Goedhart is very active on Twitter...Resources from the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's Series on Choosing... Choosing the B(right)est Fluorescent Protein Plasmids 101: Visualizing Subcellular Structures and Organelles...Organelles Resources at Addgene.org Find Fluorescent Protein Plasmids Find Plasmids for FRET Find Plasmids for...

...sections: Protocols To complement our written protocol materials, we have also created video protocols.  These...navigate various lab processes.  Our most recent protocol video detailing Streaking Plates for Single Bacterial...Check Our Our Protocol Pages Browse our Plasmid Reference Pages ... protocol video...

...Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm...below affect continuous cellular processes (e.g., protein synthesis), while amp only affects the cell wall...Kanamycin, or kan, is an aminoglycoside which inhibits protein synthesis. The kan resistance gene, NPTII, facilitates...streptomycin Spectinomycin and streptomycin both inhibit protein synthesis as their mode of action and have a very...gene provides zeocin resistance by expressing a protein that directly binds to zeocin, preventing it from...Addgene.org Addgene’s Basic Molecular Biology Protocols Over-Agar Antibiotic Plating Plasmids 101 eBook...

...adenovirus type 5 (hAdV-C5) can be considered the prototype, although many viruses infecting humans (from ... the nuclear compartment, gene expression, and protein translation. Its genome consists of “early” (E)...Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole process – such...absent, and the late genes (and their structural proteins, which would encapsidate new virions) are not ... is transient due to the fact that AdV surface proteins (especially fiber’s knob domain, and hexon), as... as well as other intracellularly-exposed proteins and downstream cellular events, all recruit strong ...J., & Wold, W. S. (1996). The adenovirus death protein (E3-11.6K) is required at very late stages of infection...

...optogenetics — and shared the protocols we use in our own labs. By 2023, we had 47 protocols and 12 guides available...experiment.     Table 1: Most popular guides, protocols, and collections of 2023. Educational Resource...Guide Optogenetics Guide γ-Retrovirus Guide Protocols Agarose Gel Electrophoresis Bacterial Transformation...around the world. Collectively, our 12 guides, 47 protocols, and 186 collection pages received over two million...a thousand words, especially when it comes to protocols. In 2013, we launched the Addgene YouTube channel...Figure 4). Four of the five most popular videos are protocol videos, where showing exactly how to execute a...

... of proteins was mutated, as reported in various studies. We made a table that listed each protein in ...then for each protein we listed all the studies that reported mutations in that protein (including how...because you could easily see how frequently each protein was mutated, you could see how big the studies ...Another example: we made a schematic of all the proteins in the class that showed the relative sizes and...sense of the similarities and differences among proteins in the class. Offer your perspective It doesn...

...antibodies against popular fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have also.... Antibodies are the most widely used class of protein-binding reagents. In the lab, their binding specificity...specificity allows scientists to target proteins of interest for labeling, quantification, purification...distribution funds our operation. Antibodies are glycoproteins that are part of the humoral immune system. ...complex antibody chips that allow assessment of many protein targets in a single sample; 3) producing recombinant...

...the target site, while Cas12a requires a PAM (Protospacer Adjacent Motif) for target cleavage. This test...test varies in the sequence of steps and the Cas protein used. Here, all the components needed for the reaction...40 min incubation. A scalable, easy-to-deploy, protocol for Cas13-based detection of SARS-CoV-2 genetic...Assay (FELUDA) is a technique that uses FnCas9, a protein that is highly sensitive to the presence of mismatches... acid detection with CRISPR nucleases. Nature Protocols 14:2986–3012 . https://doi.org/10.1038/s41596-...Wilson, M. Z. (2020). A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic...Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol for detection of COVID-19 using CRISPR diagnostics...

...primarily characterized by protein misfolding that leads to a build-up of protein aggregates termed “lewy...those that overexpress the toxic protein α-synuclein (the most common protein found in lewy bodies), have ...scientists to easily observe fluorescently labelled proteins and tissues to better assess developmental processes...modification, such as expression of fluorescent protein or disruption of a gene, can be easily visualized...quick guide to Drosophila Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources...

...companion post describing protein expression strains, where we go over the basics of protein expression in E. ...your gene, and may have even put a fluorescent protein downstream, separated by an IRES element. You did...general lab strains designed for subcloning or protein expression. If you were to include customized strains...gene Increases the stability of certain expressed proteins endA, endA1 Endonuclease I (nonspecific cleavage...fertility plasmid. A low copy-number plasmid, encodes proteins needed for bacterial conjugation. Genes listed...stringent factor Allows RNA synthesis in absence of protein synthesis Ptrc-ccdA   Propagation of ccdB-containing...

...vector to create knock-ins of fluorescent proteins, fusion proteins, or transcriptional regulators. In this...and on either side of the cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting...Find the GeneWeld vector kit CasX, a small Cas protein, is active in E. coli and human cells Two years...ago, Jennifer Doudna’s lab discovered a new Cas protein, CasX, from metagenomic analysis of microbial DNA...Cas9 to a mutated human estrogen receptor. This protein fusion stays in the cytoplasm, but with the addition...

...covalent modifications to DNA or histones, the proteins that help package DNA, were associated with gene...With the help of guide RNAs, fusions between Cas proteins and the epigenetic modifier could be targeted ...cytosine demethylation of DNA. However, several proteins in the DNA oxidation and repair pathways work ...NLS-Sa dCas9-NLS-LSD1. LSD1 is a particularly large protein, and this fusion allows for packaging into viral...exhibits increased PAM flexibility, and this fusion protein allows for LSD1 targeting to a wider set of genomic...methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nature Communications 8: . https://doi.org/10.1038...endogenous genes using programmable TALE-TET1 fusion proteins. Nature Biotechnology 31:1137–1142 . https://doi.org...

... purification or if you use a DIY purification protocol, you might have noticed that there are many options...options to elute your DNA prep. You might see protocols that recommend eluting in water, Tris-EDTA (TE...purification from E. coli is the presence of the protein endonuclease I encoded by the gene endA. Nowadays...are usually extra steps like heat inactivation, protein degradation, and/or the use of chelating agents...agents that can be introduced in the purification protocol to remove endonucleases activity. For these reasons...Check out the Molecular Biology Reference Find protocols on plasmid cloning Find plasmids for your research...

...? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead...Research Campus reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores can...Zika virus and, upon binding, produce reporter proteins. These toehold switches can be used to quickly...University, describes R-bodies, “force generating protein machines that can extend and retract over many ...different pH’s and some that are fused to fluorescent proteins. She also showed that R-bodies can rupture E. ...Casilio, a system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety...

...BBB via interactions with a similar protein called LY6C1. This protein is expressed in both BALB/cJ mice...exists to prevent substances in the blood, like proteins and viruses, from crossing into the brain. While...as the type of transgene delivered: fluorescent proteins, optogenetic sensors, CRISPR elements, etc... ... it, cross the BBB through an endothelial cell protein called LY6A (Hordeaux et al., 2019; Huang et al...capsids that rely on interactions with those two proteins to cross the BBB are often not effective in NHPs...in rats. For more information on reagents and protocols, visit https://clover.caltech.edu. Figure adapted... selected for its ability to bind to the LY6C1 protein in order to reduce strain-specific tropism in mice...