...a programmable RNA-binding protein, creating the fluorescent fusion protein dLwaCas13a-NF for in vivo ...has many applications. That said, SpCas9 is not the only game in town, and other Cas proteins like SaCas9...form of “bait” to identify new CRISPR-associated proteins within the bacterial genome. From this analysis...categories based on the architecture of the CRISPR protein in question: C2c1, C2c2 and C2c3 (short for Class...consistent with known roles for HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a... the sequence of interest and an inactivated fluorescent RNA reporter. If the target sequence is present...resulting in activation of the fluorophore. The fluorescent signal can be used as an indicator to determine...

...Developments in light sheet fluorescence microscopy Modern light sheet fluorescence microscopy was first pioneered...such as whole mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories... advances in microscopy has been light sheet fluorescence microscopy (LSFM). The underlying method was...colleagues and originally named orthogonal-plane fluorescence optical sectioning (OPFOS) (2). Arne Voie, David...laser beam into a thin sheet to illuminate a fluorescent sample and captured the reflected light using...configurations’ various features see (4,7).   Light sheet fluorescence microscopy vs. confocal microscopy In confocal...developed AAV (adeno-associated virus)-based fluorescent labeling methods that can be used with 3DISCO...

...integration of fluorescent tags in neurons. Their toolkit named ORANGE (Open Resource for the Application of Neuronal...engineered SECURE adenine base editor variants using a protein-truncation strategy with reduced RNA-recognition...abilities of C-to-T and A-to-G base editors into one protein. They fused an activation-induced cytidine deaminase...Editing) mediates targeted genomic integration of fluorescent tags in neurons.  Read the paper in bioRxiv ...with a com aptamer which binds an aptamer binding protein for packaging into a lentivirus-like particle. ...tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine deaminase for RNA editing...bioRxiv Find the ORANGE toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle...

...to change protein levels? Until recently, no such tool existed for rapid, customizable protein degradation...both short-and long-lived proteins, and it can even distinguish between protein variants when used with ...addition of exogenous TRIM21 protein along with an antibody targeting the protein of interest led to rapid...has deposited plasmids encoding fluorescent reporters and other proteins controlled by GAL4 and ZIF268....James and Schuh labs. Clift et al. applied TRIM-away to nine proteins in ten ddifferent cell types, including...Plasmids Podcast! TRIM-Away: Targeted endogenous protein degradation Article contributed by Mary Gearing... cell populations, co-microinjection of TRIM21 protein and the target antibody works well. In larger cell...

...interest (or, if your protein is fused with a fluorescent protein), fluorescence microscopy can theoretically...Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors... place to screen your clones. Depending on the protein, we have used Western blots, dot blots, ELISAs,...population of transfected clones. The technique is applicable to a wide variety of cell lines, from HEK to ...

...co-localization immunofluorescence, cell-type specificity. Viruses that only express the fluorescent proteins are ...Thankfully the Deisseroth constructs coexpress a fluorescent protein, which allows for easy identification of ...vectors for anterograde axonal tracing with fluorescent proteins in nontransgenic and cre driver mice. Current...optogenetics plasmids! Find AAV resources! Find fluorescent protein resources!    ...general rule of thumb, if you can detect a strong fluorescent signal (without antibody manipulation), then ...22430017. 2. Fenno L, et al. The development and application of optogenetics. Annual review of neuroscience...PMC3225502. 4. Mattis J, et al. Principles for applying optogenetic tools derived from direct comparative...

...Western blots are a great tool to identify a protein of interest in a complicated solution like cell ...— and what if you want to detect more than one protein in your sample? Or what if something weird happened...membrane, which is sturdier and retains the blotted protein better than a nitrocellulose membrane. If you have...probe for. Even using PVDF, some of your target protein will be removed from the membrane along with the...methods. If you are planning to detect multiple proteins through several rounds of stripping and reprobing...your signal by starting with the least-abundant protein and working toward the most abundant. If the abundance...to preserve your sample. Pro tip! Because some protein is lost each time you strip a blot, you should ...

...feature inaccurately. The best example is fluorescent proteins, which often come in closely related versions...misidentification, we augment the database with new fluorescent protein variants. The future of DNA feature identification...system was enhanced to allow searches for a perfect protein sequence match even when the DNA sequence has changed...Collections”, which are shareable databases of DNA and protein sequences, the next step is to enhance Collections.... We are working on a way for SnapGene users to apply features from a reference strain to newly sequenced...

...single-domain antibodies genetically fused to fluorescent proteins and used to visualize target antigens in...around using sdAbs in diagnostic applications. For example, fluorescently and radioactively labeled sdAbs...Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids encoding...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative...inspired this versatile family of tools, the applications they have already enabled, and the discoveries...

...method for identifying protein-protein associations. This method uses a bait protein fused to a biotin ligase...PrM N and C   Detecting protein-protein interactions with DULIP Proteins rarely act alone. Molecular...which “bait” proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with...with Rosella, a pH-sensitive fluorescent biosensor A pH-sensitive fluorescent biosensor-based assay system...ligase to biotinylate proximate proteins in the cell. These candidate proteins can be enriched via biotin ...of fusion proteins, requires less biotin, and shows enhanced labeling of proximate proteins. Overall BioID2...BioID2 improves the efficiency of screening for protein–protein interactions. The Roux lab has deposited HA...

...will not produce the fluorescent protein while areas that are exposed will fluoresce. With these images,...transcriptional regulator Their system is based on the AraC protein that controls transcription from the PBAD promoter...reached the level of expression needed for most applications.   Figure 1: With blue light, the VVD...

...categories: antibiotic/herbicide resistance and fluorescent proteins. Selection using resistance genes usually...would only then kill the desired plants! PCR or fluorescent markers are currently used in these situations...could be achieved much easier than by PCR or a fluorescent microscope. First, red CRISPR-positive transformants...culture), which is great for a wide variety of applications. But some techniques require you to select against...

...Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] and Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target distinct...the SARS-CoV-2 nucelocapsid protein, one of the virus’s most abundant proteins and a critical component ...SARS-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M]...transcriptase enzymes, inhibition of host cell proteins, and modifications to the pegRNA. The Britt Adamson... factors impacting prime editing and found the protein La (a small RNA-binding exonuclease protection ...prime editing with an endogenous small RNA-binding protein. Nature. 2024 Apr 3. doi: https://doi.org/10.1038...immunoblotted against Anti-SARS-CoV-2 Nucleocapsid Protein antibodies #211756 (left panel) or #211757 (middle...

... art have a look at our Microbiology and Fluorescent Protein Resources and get started!  I learned a lot...paper thin and very portable microscope. Less fluorescence admittedly, but still so beautiful! Enabling...cellular imaging with ultrasound. In his research he applies naturally occurring bacterial “molecular buoyancy...biomolecule-based dyes. This technology is widely applicable as bacterial expression of dyes allows dye visualization...

...constitutively express a red, green, or blue (RGB) fluorescent protein (4a-c). When high doses of this viral cocktail...are easy to trace and not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color...A cocktail of three AAV vectors expressing RGB proteins under the control of an inducer, i.e. tet-off ...morphology studies when used as part of a two part fluorescent tag-then-express system that allows for controlled...morphologic studies), as well as translational applications (mechanistic studies and gene therapy treatments...

...macromolecules used in scientific research such as DNA or protein. It comes as no surprise then that stability of...crRNA (tracrRNA) which interacts with the Cas9 protein. These RNA molecules are vulnerable to attack by...gRNA is surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same...simple LED light. Lighting up your gRNA with fluorescent dyes In a scientific world where it often seems...seems like everything glows, why not spread the fluorescence to your gRNAs? Dye labeling the RNA in your ...were developed before CRISPR-Cas9, for use in applications like siRNA and antisense oligonucleotides. Some...How to design your CRISPR gRNA CRISPR 101: Ribonucleoprotein (RNP) Delivery Components of CRISPR/Cas9 ...

...Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells   ...and many more!...tools useful to any molecular biologist Tips for protein purification Replica plating Anti-CRISPR Retroviral... Molecular biologist with Biogen interested in applying CRISPR for mouse genome editing. Post: Generating...

...Since the anti-GFP nanobody is able to bind fluorescent proteins derived from A. victoria the functionalized...locus and labels the proteins in its vicinity with biotin. The biotin-labeled proteins can then be isolated...investigated protein binding at the TERT gene, but this method enables the unbiased discovery of proteins involved... proteomics: developing and applying new technologies to quantify proteins in tissues, cells, and biofluids...nanobodies described here can be used to target any protein fused with GFP, YFP, mCerulean, or other related...: 29915061.        A new method to identify proteins occupying a specific genomic locus: Caspex  Article...group, developed a CRISPR-based approach to detect proteins that associate with specific genomic loci. He ...

...polymerase, T7 polymerase, EcoRI, and various fluorescent proteins. Dr. Duedu’s plans for these plasmids have...bioinformatic and proteomic tools to discover genes and proteins associated with pathogenesis. In his future work...concluded, Addgene and Seeding Labs will be opening applications for a second round of plasmid awards on June...excited to discover what new and creative ideas applicants will propose for plasmids from the Addgene community...regions in the near future. Additional Resources Apply to Recieve Equipment through Seeding Labs' Instrumental...

...identification of plant compounds and insecticidal proteins to the development of renewable biotechnology ...system pDawn to clone and express newly identified proteins and enzymes. These compounds will later get tested... nutritionally and therapeutically significant proteins. In this project she aims to create transgenic...desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc.), Nong Lam University, Vietnam...and analysing the insecticidal potential of Cry proteins in order to manage insect pests and to support...this project the expression of isolated bacterial proteins can be achieved using a bacterial expression system...vector and the pOKD plasmids. The recombinant Cry proteins will then be screened for toxicity against local...