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...colleagues devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers”...are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...fluorescent sensor for real-time measurement of extracellular oxytocin dynamics in the brain. Nat Methods....Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific RTK activation in...biosensors for monitoring RTK activation by shift in cellular localization. Image adapted from Farahani et al...so this sensor will be a valuable tool to study cellular physiology and metabolism under various conditions...biosensors, and its affinity is better-matched to cellular concentrations of lactate. The microfluidic screening...

...beta subunits. Both subunits have short intracellular ends and large ectodomains containing ligand-binding...signaling pathways and the linkage of intra- and extracellular environments. Found on nearly every cell in ...unique antibodies that bind discrete subunits or particular subunit pairs. Some of the antibodies in the ...the cell membrane, physically connecting the extracellular environment and the cytoskeleton. Most integrins...affinity for ligands. When integrins interact with extracellular ligands, their ectodomains can extend and open...mediating cell adhesion and migration, initiating intracellular signal cascades, and allowing agile responses...it is resisted by the ligand embedded in the extracellular environment. This further stabilizes the extended...

...federal regulators. But the speed is impressive on a larger scale, too. The first paper reporting genome editing...later, an A→G base editor (ABE) followed based on similar principles (Gaudelli et al., 2017). These editors...would be tested for their base-editing efficacy in cellular models of KJ’s CPS1 mutation. Every genetic target...base editors that could target KJ’s mutation in cellular models tested both the SpG and SpRY variants of... of Cas9. With many PAMs available, a large number of gRNAs could be screened for the most efficient editing...PAM preferences (Silverstein et al., 2025). In cellular models of KJ’s mutation, SpG Cas9 + ABE8e performed...Up to 5 bystander adenine bases were edited in cellular tests, in addition to the target base. However...

...of polylinker sequences of molecular vectors". Proceedings of the Belarusian State University Series of...for allelic exchange Read our molecular biology reference Watch molecular biology protocol videos ...contain sacB and will not grow on sucrose. One popular counterselection vector is the pDS132 sacB-based...counter-selection. Anecdotally, there is also a large amount of user-to-user variability, and in some ...colonies turn subtly blue or magenta. This is particularly valuable when first optimizing conjugation in...modification of bacterial genomes. It can accomplish large deletions, insertions, and even precise point mutations...Harvard. Jake received his MD and a PhD in Cell and Molecular Biology from the University of Pennsylvania, and...

...binding partner avidin are commonly used today in molecular biology for an array of different techniques and... interaction and the uses of biotinylation in molecular biology! Learn about in vivo biotinylation of ...biotin is most commonly known for its role in molecular biology, biotin (Vitamin H or Vitamin B7) is first...carboxyl groups to organic acids to produce important cellular metabolites. Biotin acts as a carboxyl carrier...temperature or organic solvents. Uses of biotin in molecular biology Biotin is a relatively small, water soluble...biotin and its strong interaction with avidin for molecular biology and biotechnology purposes.  Once a polypeptide... or co-expressed with avidin or a structurally similar protein that was discovered in the 1960s, strepavidin...

...this post: Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized genetic screening technique...editing Anti-FLAG Frankenbodies for modular imaging of cellular targets by: Mike Lacy Adding to their... a set of anti-FLAG frankenbodies, a flexible, modular tool for labeling. Here, Liu et al. engineer a ... cells, Western blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to...single expression cassette is advantageous to Cas9’s larger expression requirements (Figure 1), but Cas12a ...create a genome-wide library capable of performing similarly to Cas9 screens in a compacted library. Overall...genome regulation and editing by: Rachel Leeson The large size of CRISPR-Cas effectors and fusion proteins...

...lesser-known Cas enzymes while also highlighting their similarities and differences to Cas9. DNA targeting Cas...genome edits, but it is effective at wiping out large regions of DNA and has anti-viral application potential...facts – PAM requirement: AAG or TTC  Best for: very large deletions.   Cas12a – The staggered cutter  Originally...Cas12a cleavage results in 5 bp overhangs and is particularly convenient for targeting experiments requiring... even a single mismatch in the target sequence Similar to Cas12, it is also significantly smaller in size... size than traditional Cas9. Another similarity to Cas12a is that Cas14 also indiscriminately cleaves ...it targets RNA instead of DNA. This makes it particularly useful in generating transient edits of signaling...

...McKenna, Ph.D., is Associate Professor of Molecular and Cellular Biology at Baylor College of Medicine and...Professor and Chair of the Department of Molecular and Cellular Biology, and Editor-In-Chief of the Open...understanding of how these processes are regulated at the cellular, tissue and organ level by nuclear receptors (... receptors and their ligands NRs represent the largest family of transcription factors found in metazoans... organs because of their established roles as molecular sensors and governors of endocrine-hormone signaling..., including hypertension, cancer, diabetes, cardiovascular disease, cholesterol gallstone disease, senescent...provided clues to their previously uncharacterized cellular roles. In the current phase of NURSA we are seeking...

...their detection can suffer from photobleaching and cellular autofluorescence (Tsien, 1998). What sets HiBiT...activity is compatible with both intra- and extracellular environments, making it effective for analyzing...from the Promega Plasmid Collection that express similar constructs available at Addgene. Read the original...HiBiT-TP53 Fusion Vector at Addgene! Following extracellular RNA editors Tracking factors that utilize exosomal... hypothesized that if ADAR1 were carried by extracellular vesicles (EVs), edited transcripts could endow...endoplasmic reticulum quality control navigate a large and interconnected system of dynamic and redundant... endocytic trafficking in real time through extracellular-specific and full lytic measurements. Find CFTR-HiBiT...

...the new fragment thereby generating a complete, circular plasmid that can be easily maintained in a variety...companies now sell fast digest enzymes that can digest large amounts of DNA in as little as 10 minutes, but check...will need to use a phosphatase to prevent re-circularization of the backbone plasmid (see below). You should..., the DNA migrates toward the anode (+ charge). Larger fragments of linearized DNA migrate slower than... to anneal to one another and form a complete, circular plasmid. You then add DNA ligase to covalently...in the transformation due to background re-circularization and contamination from uncut plasmid. 4. Transformation...Additionally, if your final product is going to be very large (>10kb) you might want to use electro-competent ...