...dTomato) that promote interactions between the FPs interfere with native interactions, preventing proper ...the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity and...higher order aggregates. This property can be tolerated in some applications (e.g. labeling of cells or...homodimers or higher order oligomers can be analyzed by several techniques. Gel filtration and SDS-PAGE under non-denaturing...reflect the likelihood that fluorescent proteins interact or oligomerize in cells? This question is largely...fluorescent protein’s propensity to homodimerize. Several studies have used the OSER assay to demonstrate...

...Performance dCas9-VP64 activation is generally thought of as the “first generation” CRISPR activator. While it ...initially published in 2013 (Bikard et al., 2013, Perez-Pinera et al., 2013). In the years that followed, innovative...Performance SunTag performs better than first generation activators while showing lower activation levels...to recruit transcription factors. Performance Generally, VPR is found to have significantly higher activation...method, dCas9 is fused to CBP, an histone acetyltransferase domain capable of rearranging chromatin structure...588 . https://doi.org/10.1038/nature14136 Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein...

...your study is aimed at developing human therapies. If a therapeutic is the end goal, it is best to work with...Targeting is also important in the development of therapies when expression must be restricted to specific... crossing the blood-brain barrier.  Another consideration when picking ligands for chemogenetic studies...ligands. However, they retain their intracellular interactions and still set off the same pathways, giving ... (Saloman, JL, et al., 2016).   DREADD ligands Several DREADDs, including hM3D(Gq), have been derived ...& Gold, M. S. (2016). Gi-DREADD Expression in Peripheral Nerves Produces Ligand-Dependent Analgesia, as...

... In this article we will discuss general viral safety considerations and best practices for handling lab-grown... basic biological research as well as clinical therapy. These biological tools are useful because they... separate plasmid (envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein... and the same holds true for types of viruses generated in the lab. The four most common types of viruses... lab and require additional safety measures.   General viral handling safety   Containment and personal...resistant to ethanol and isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some...

...specific to the acceptor In order for FRET to occur several constraints must be met. The donor and acceptor...appropriately for energy transfer via dipole-dipole interaction. The efficiency of FRET can be measured for a...FRET range, FRET efficiency is used to infer an interaction between the fluorophores and serves as a small-scale...As the choice of fluorophore pairs is a key consideration for a FRET experiment, the possibilities can...of fluorescent proteins available. Fortunately, several excellent reviews have collated data from various... typically used for studying protein-protein interactions by measuring the change in FRET efficiency, ..., FRET experiments can probe protein-protein interactions, measure the concentration or activity of small...

...circuits with functional relevance following a set hierarchy. MoClo originally developed from Golden Gate Cloning...organization schema MoClo can be broken down into three hierarchical levels: individual DNA parts, transcriptional...For example: in a cell there are proteins which interact with other proteins and with DNA, and these complexes...are multigene systems whose protein products can interact with the DNA that encodes the protein itself or... or other genes within the circuit. These interactions yield a cellular phenotype, translational output...can support a larger cellular engineering goal, several of which are described below. Multigene circuits...promoter for a given peptide can easily be tested by generating Level 1 TUs with unique promoters, and the ideal...

...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...churn out antibodies against the injected protein. Several weeks later, blood from the animal is drawn and...antigen binding, the variable regions alone can be generated as a fusion protein. An ScFv is made up of the...your samples to a standard curve. This curve is generated using a serial dilution of a known amount of protein...for other experiments to study protein-protein interactions. Flow cytometry Flow cytometry is used to detect...2014.   In vivo uses For both research and therapeutic purposes, antibodies can be used to affect cellular...functional grade antibodies. Some of the research and therapeutic uses overlap: antibodies can bind to and inhibit...

... Overview of the CRISPR classification hierarchy. The hierarchy flows from Class to type to subtype to...the array of CRISPR technologies. Learning the general principles of this classification system will help...classifications were placed into a fairly simple hierarchy: class to type to subtype to variant. Despite ...seemingly complicated steps to arrive at this hierarchy, the main aspects of each class and type are easy...Class 1, and are the most common type of CRISPR overall. Type I systems degrade large sections of DNA thanks...most common. Figure 4: Overview of the general CRISPR type VI mechanism. The Cas13 effector binds...diversity of rare CRISPR-Cas systems with deep terascale clustering. Science, 382(6673). https://doi.org...

... virus may acquire mutations to escape these therapeutics. Jesse Bloom’s lab at the Fred Hutchinson Cancer... Figure 1. (A) SARS-CoV-2 RBD DMS Library generation and sequencing. From Starr et al Cell., 2020....synthetase complex, the team investigated whether the generated antibodies could bind these aaRSs. They found ... Gräslund lab plans to continue their work by generating antibodies for additional aminoacyl tRNA synthetases...nanoparticle can protect proteins from unfavorable interactions in the cell, improving biosynthetic pathway ...represent 0.001% of the population.  Because indel generation by Cas9 can occur at a steady rate, scientists... Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes AAV1 and...

...Evan’s team generated their Myc antibodies. At first, anti-c-Myc [9E10] was just one of several new tools...over the years, several alternative antibodies against the Myc tag have been generated in an attempt to.... However, it was (and still is) infeasible to generate highly specific antibodies for every potential...would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different synthetic...epitope within that region. You’ll notice that “generate a reliable antibody for protein tagging approaches...