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...recently developed a photosensitizer composed of the protein complex complex and the MG-2I-dL5** fluorogen that...thus forming the full FAP-TAP complex (MG-2I-dL5**).   A FAP-TAP complex will light up if excited with...cells. The photosensitizer used in this work is composed of two parts: 1) an activatable protein (the FAP...-TAP MG-2I-dL5** MG-2I-dL5** is a new FAP-TAP complex where dL5** is the FAP. It’s job is to bind MG-...fluorogenic dye.  MG-2I is the TAP portion of the complex and is derived from the malachite green (MG) dye...modified with the addition of two iodine residues. Compared to MG, MG-2I has increased ROS generation when...with these fluorescent proteins. He et al also compared MG-2I-dL5** head-to-head with KillerRed, a genetically...

...many opportunities to learn from other biotech companies at workshops and can use what we learn to improve...office space shared by numerous emerging biotech companies. The space was modern with clean bold colors of...hallway for people to take a break and relax. Companies at LabCentral get access to a variety of experimental...how LabCentral aims to enable emerging biotech companies, I was excited to go there because both LabCentral...must first fragment a DNA sample into many small components. Without proper processing, these small DNA strands... workshop participants from different biotech companies to interact and learn more about one another. ...expand my skills and network with other technicians/company representatives. I got to talk with a technician...

...discovery in the 1990s and quickly rose to popularity. Compared to CRISPR, RNAi methods are often quicker and ...create pri-miRNA, which is then processed by a complex called Drosha to pre-miRNA. miRNA plasmids are ...RNA duplexes bind to the RNA-induced silencing complex (RISC). After binding to RISC, the RNA duplex is... can simply block translation machinery. The composition of the RISC varies, but at the very minimum requires...recruits other silencing proteins. Figure 1: Comparison of shRNA and miRNA processing and silencing pathways...typically range from 19–22 nucleotides and are 100% complementary to their target sequence. However, you can’t... day. You need to include both sides of the complementary RNA duplex and the hairpin loop sequence (Figure...

...isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds and inactivates the lac ...to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional enzyme...enzyme. This process is called α-complementation. The system described above was put to practical use in the... inserted it into a plasmid, creating an α-complementation cloning vector. When a cloning reaction goes...interrupted as shown in cell A, and thus will not complement the cell's β-galactosidase mutation. An unsuccessful...functional β-galactosidase enzyme through α-complementation (cell B). As shown on the representative plate... use a proper plasmid (i.e. contains the α-complementation cloning MCS): pGEM-T, pUC18 and pUC19, and ...

...cerevisiae yes - This can complement ura4- S. pombe, but the complementation is weak. yes   LYS2 L-lysine...vector propagation. This post, along with a future companion post on mammalian vectors, will catch you up on...vectors? Yeast are eukaryotes and thus contain complex internal cell structures similar to those of plants...in the lab typically contain all the necessary components to allow shuttling between E. coli and yeast ...an organism to synthesize a particular organic compound required for its growth. Many auxotrophic strains... including a copy of a functional gene which complements the host’s auxotrophy. When grown on media NOT...LEU2 L-leucine no S. cerevisiae yes - This can complement leu1- S. pombe, but multiple copies are required...

...of the protein MLH1, a component of the MutSα–MutLα mismatch repair (MMR) complex, to temporarily inhibit...45-fold) higher across different genomic sites compared to PE1 (Anzalone et al., 2019). PE3: Resolving...recognized as an MMR intermediate, and the MutSα–MutLα complex may either selectively excise the edited DNA flap...the unedited strand. Temporarily inhibiting this complex may allow time for 5′ flap exonucleases and DNA...nuclease activity (Chen et al., 2021). In a system as complex as prime editing, every efficiency counts! The ...deliver with AAVs but still efficient at long and complex edits. The team also developed PE6e-g editors with... the truncated pegRNA can still guide Cas9 and compete with full pegRNAs for access to the target site...

...projects in introductory labs. Genome projects use computational methods to identify genes by their similarities...abnormalities due to the deletion (i.e. the plasmid complements the deletion). If the phenotype is corrected,..., well-studied and inexpensive. In addition, a complete set of well-annotated deletion strains is available... the absence of methionine (i.e. the plasmid complements the deletion). Find the Plasmids Here Plasmid...blotting in other experiments (Figure 2).   Complementation reveals conservation of function Students have...Addgene, to successfully analyze Met protein complementation. In the procedure diagrammed in Figure 3, students...promoter controls MET/Met gene expression, complementation should only occur when galactose replaces glucose...

... for a new generation of ABEs), consists of 3 components:  a deaminase (TadA, originally from E.coli,... or Cas nickase)  a guide RNA that targets the complex of TadA and dCas to the genomic DNA of interest...effect of removing the wild-type TadA from the complex the researchers developed a second set: The ABE8...resulted altogether in 40 new ABE8 variants. When comparing on-target DNA editing efficiency in a human cell...found that the smaller version, ABE8.x-m, showed comparable editing behavior to ABE8.x-d. Overall, ABE8s ...showed a median increase of 1.94x editing activity compared to ABE7.10. Depending on the position in the protospacer...line while reducing indel frequency more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase...

... human body is made up of organs, cells comprise compartments and structures, called organelles. Take ... the subcellular localization and composition of cellular complexes are important steps in understanding...that the fusion protein localizes properly by comparing the signal from the fusion itself to the wild-...and potentially which other proteins it forms complexes with. These “colocalization” studies give insights...within the same subcellular structure or protein complex. By coexpressing both the marker protein and the...possible to determine the location of proteins within complex structures. In order to interpret colocalization...quantifiable results and statistics that can be used to compare between different experimental setups. A colocalization...

...contains a composite of GenBank, EMBL, DDBJ, and PDB sequences and may be the most comprehensive for searching... Tool (BLAST) provided by NCBI. BLAST helps us compare the sequencing results of the plasmids in our repository...deletions, or insertions. We use blastp or blastx to compare our sequencing results to protein sequences to ...page, the first decision to make is whether to compare a Sanger sequencing result to a single known reference...BLAST search to perform and will save you time compared to other types of BLAST searches. If you do not...megablast, but can return longer alignments to compare with your sequencing trace file. Unlike megablast...consistently run our blastx searches with the “Low complexity regions” filter unchecked so that these regions...

... most successful computer languages are free to use. Of the 15 most popular computer programming languages...engineered DNA systems capable of storing data and computing inside living cells. He's also a co-founder of...digital genetic switches that enable cellular computing. These logic gates built from transcriptors are...are available at Addgene as a BIOFAB kit comprised of well-characterized bacterial transcription and translation.... What the BioBricks Foundation is seeking to accomplish is to make the engineering of biology much easier...carefully engineered to support reliable functional composition. In other words, these parts can be used over...logic shows up in mechanical systems, plumbing, computers, and now through BIL gates we've implemented them...

... as the building block of our understanding of complex biological processes. Many tools exist to perturb...silencing,” of any gene that contains sequence complementary to the dsRNA (Wilson and Doudna, 2013). RNAi... translation, or chromatin modifications of complementary gene sequences (Figure 1). As a result, gene...unmodified dsRNA elicits immunogenic reactions that complicate the effectiveness and interpretation of RNAi.... of interfering RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has...Stainier, 2020). Additionally, a study that directly compared CRISPR to RNAi in the context of a high throughput...data generated from both methods did a more comprehensive gene list emerge that accurately reflected the...

...scientific art competitions where creations can go on display for all to enjoy. The largest competition of this... University’s Annual Art in Science competition. This competition has gained so much public interest for...moonlighting as artists can compete in the Nikon Small World Photomicrography Competition - a 40 year old international...voyage. Previous destructive techniques were incompatible with the need to conserve these unique samples...been featured in Time magazine . Other smaller competitions are now taking place and gathering momentum,...international competition that last year received over 2,000 entries. Do you mix your passion for art and...

...them for use in your lab. Structural Comparison Functional Comparison Advantages of Nanobodies How to Use...for a comparison of nanobody, HCab, and traditional IgG antibody structures. Functional comparison Strong...nanobodies and how their structure and function compare to IgG antibodies, as well as how to produce them...Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, nanobodies are smaller ...incubated together without forming large oligomeric complexes. Antibodies are also too large to be expressed... antibody incubation. See figure 4A below for comparison of this one-step staining with nanobodies vs ...localization patterns as cells stained for one target (compare nanobodies staining in figure 4A vs 4B). How to...

... function as a kind of “GPS for the genome” to compute coordinates for genetic elements like protein, ...action at a distance.” Through the marvel of DNA compaction, an enhancer can increase the expression of a...millions of base pairs away. (For details on DNA compaction and the structural proteins that make it possible...appendix DNA is a 3D fractal). Enhancer biology is a complex and dynamic field. We’re going to focus on a tried...details. DNA is a 3D Fractal Figure 2: DNA compaction. Graphics courtesy of beyondthedish (left) and...People are wrong. In reality, cellular DNA is a complex three-dimensional globule of chromatin. Chromatin...tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes per gram—and form...

...But what exactly is an antibody, and how is a component of the immune system useful as a research reagent...heavy chains, and the rest of the Fab regions are completed with the Ig light chains. The variable region ...production strategy has advantages and disadvantages compared to each other. Polyclonal antibodies These antibodies...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. Because... their lower affinity for their target protein compared to whole antibodies (Crivianu-Gaita 2016). Get...Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies are the...fluorescence. Western blots are frequently used to compare relative levels of protein expression between cell...

...derived from the more complex type III-A and III-B systems. Though it was composed of many protein subunits...might be phylogenetically classified as the more complex Class I systems that contain multiple subunits,...evolution, were also working on the project. Using computational observations, Koonin and Makarova found that...the domains into a single protein, shedding the complexity of its ancestors. They classified the system ...system contains two pieces: a single protein, computationally described as a fusion of four Cas7 proteins...significantly reduced amount of off-target toxicity compared to other RNA editing methods — streamlining experimental...mammalian cell lines. However, in those same comparisons, the AbuGoot lab reported no detectable effect...

... advise pharmaceutical companies, biotech start-ups, health insurance companies, and governments on the...the part of the company involved: Research & Development (R&D) teams at pharma companies carry out preclinical... acquisitions of or collaborations with other companies. Medical Affairs groups work on building awareness... patients, shaping discussions with insurance companies about the pricing and coverage of drugs, and developing... teams, reflecting the need for all parts of a company to collaborate whenever it faces an important challenge...investments over the next 3, 5, or 10 years. Complicating these challenges is the fact that they occur...their jobs most, get pleasure from taking up such complex challenges, quickly breaking them up into smaller...

...Table 1: Taking it a Step Further - Big Company v. Small Company Big (e.g. Pharma) Small (e.g. Biotech...advancement in one company, good performers in for the long haul Limited movement in one company, likely will...academic silo to tenured professor.  Non-academia encompasses millions of choices including pharma, tech transfer...options in nonprofit science. No one list can ever encompass them all. We can’t designate non-academic jobs...and teach students Collaborations may be less “complicated” - for example, easy to initiate a collaboration...More sources of training and resources within the company Must find own sources of knowledge often externally...every 3-6 years—a short and exciting ride Complicated matrix structures are common Small integrated...

...protein. Proteins located in membrane-bound subcompartments, like the mitochondria or nucleus, will likely...loading buffers, as most Bradford assays are not compatible with detergents like SDS, while BCA assays are...their differing chemistries. You therefore cannot compare Bis and Tris gels or blots to each other. Once ...protein-based blocker is either degraded or not completely mixed together.   Commercially available chemical-based...lower cost, and smaller size. However, HRP is not compatible with sodium azide, a common microbial agent used...phosphates have a linear reaction rate and are compatible with antimicrobial agents. Chemiluminescence ...chemiluminescence enzymes, they are more likely to be compatible with stripping and probing.   Imaging At the ...