We narrowed to 371 results for: Fus
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TypeBlog Post...from the Zhang Lab. It's name, though a little confusing to the uninitiated, contains much of the information...directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene through a 2A peptide...the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at its N-terminus. This tag can...
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A new platform for fast functional characterization of potentially cancerous mutations
TypeBlog Post...they identified included gene amplifications and fusions, missense mutations, and indels (Ng et al., 2018...carrying a single gene with a defined mutation or gene fusion. Plasmids carrying the corresponding wild-type ...Mutations in Cancer.” Of all the mutations and fusions represented in this collection, approximately 80%... -
What's New in CRISPR - September 2019
TypeBlog Post...genome editing by fusing Cas9 to a mutated human estrogen receptor. This protein fusion stays in the cytoplasm...vector to create knock-ins of fluorescent proteins, fusion proteins, or transcriptional regulators. In this... -
CUT&RUN: An Improved Method for Studying Protein-DNA Interactions
TypeBlog Post...by using the DNA cutting activity of a Protein A fused micrococcal nuclease (MNase) to specifically isolate... against the protein of interest. The Protein A fused MNase is then added and Protein A binds Immunoglobulin...protein interactions Learn about other types of fusion proteins Study chromatin interactions with CAPTURE... -
Fluorescent Proteins 101: Fluorescent Biosensors
TypeBlog Post...biosensors contain a single fluorescent protein fused to a domain that senses the concentration of a given...an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin kinase... an intracellular or extracellular molecule? By fusing a biosensor to an organelle-specific targeting ... -
Early Career Researcher Toolbox: Free Online Molecular Biology Tools
TypeBlog Post... for Goldengate cloning Design primers for In-Fusion Cloning Design primers for Gibson Assembly and ...restriction enzyme cloning, Gibson assembly, or In-Fusion cloning. NEB Ligation Calculator: This tool works... from five different insert:backbone ratios. In-Fusion cloning molar ratio calculators Lab math calculators... -
Plasmids 101: Gene, enhancer, and promoter trapping
TypeBlog Post...an exon of a gene to generate a transcriptional fusion. This may disrupt the endogenous function of the...the gene product resulting in a transcriptional fusion, much like promoter traps. While this system does...disrupt the gene it monitors due to transcriptional fusion. Examples of promoter, enhancer, and gene... -
Choosing Your Perfect Empty Backbone
TypeBlog Post...available – you can always add an epitope tag or a fusion protein. Mammalian vectors One of our most popular... cells. The addition of an in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo... offer a collection of vectors carrying several fusion proteins. Pick your promoter, selection cassette... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post...sites to tether many copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors...to the plasma membrane; this sequence prevents diffusion of the mCherry labeled mRNA and keeps it in a ...SunTag, a synthetic scaffold that can recruit GFP fused to the SunTag-specific antibody scFv. Each time ... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog Post...conditionally and reversibly degrade proteins of interest fused to a degron, a domain that induces degradation. ...In addition to the engineered SpCas9, they also fused the SpCas9-NG with the activation-induced cytidine...light-activated-dynamic-looping (LADL) system. The group fused dCas9 to a truncated CIB1 protein from Arabidopsis... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog Post...binding protein known as GltI. When the resulting fusion protein binds to glutamate, conformational changes...original iGluSnFR in the pMinDisplay backbone is fused to a specific peptide segment (PDGFR TMD) that anchors...Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane domain in the mammalian... -
Targeted Mutagenesis with EvolvR
TypeBlog Post...diversification by targeting a site of interest with dCas9 fused to the Activation Induced Cytidine Deaminase (AID...two key components: the error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis...consists of an error-prone polymerase (PolI) that is fused to a nicking Cas9. A guide directs Cas9 to a site... -
Hot Plasmids - February 2022
TypeBlog Post... mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP,...Leeson The large size of CRISPR-Cas effectors and fusion proteins, such as Cas9 and Cas12a, creates challenges...from Xiaoshu, et. al. 2021. Created by the fusion of Cas12f with a transcriptional activator, CasMINI... -
Antibodies 101: Monoclonal Antibodies
TypeBlog Post...producing monoclonal antibodies is from a hybridoma, a fusion between an antibody-producing B cell and a myeloma...cells from an immunized animal, those B cells are fused to myeloma cells at a 1:1 ratio and then cultured...medium called HAT. This medium selects against unfused myeloma cells and antibody-producing B cells that... -
GPCRs: How Do They Work and How Do We Study Them?
TypeBlog Post...beta-arrestin-TEV protease fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription... et al., 2017). In this technique, the receptor fused to APEX2 catalyzes the biotinylation of proteins... -
Plasmid Tools for Microbiome Studies
TypeBlog Post...proteins into one particle within the cell, the lab fused the red fluorescent protein (RFP) to a self-assembling...microcompartment protein. The expression of this fusion is under control of the arabinose-inducible promoter... with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are washed to stop the production... -
Plasmids 101: The protein expression toolbox
TypeBlog Post...interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor to your gene of interest...activators (CRISPRa) are transcriptional activators fused to a dead Cas9. The system is led, via a custom ...specific ligand to mark the tag and whatever it is fused to for degradation. Since the tagged protein of ... -
Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP
TypeBlog Post...showed that nanobodies could be fused to other proteins, and these fusions retained the ability to bind ...Caussinus, E., Kanca, O., and Affolter, M. “Fluorescent fusion protein knockout mediated by anti-GFP nanobody.... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog Post...our FRET Resource page and can be used to create fusions with a gene of interest. These plasmids will be..., where the donor and acceptor fluorophores are fused to two separate proteins. (In contrast, intramolecular...achieve, because the ratio of acceptor to donor fusions varies with transfection efficiency and any unpaired... -
Hot Plasmids February 2024
TypeBlog Post...with Peptide-Assisted Genome Editing (PAGE). They fused the Cas enzyme to cell-penetrating peptides and ...guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals... et al. (2023). StayGold variants for molecular fusion and membrane-targeting applications. Nature Methods...