We narrowed to 641 results for: UTY

... their PhD and postdoctoral training. Addgene executive director Joanne Kamens has written a 5-part blog...pursue leadership opportunities at the local, institutional, regional, and national levels. These skills... But how does one gain such experience? Your institution may offer occasional workshops on leadership ...Harvard Medical School and then Boston University (institutions where I’ve worked as a postdoc). I soon realized...that many of the issues faced by postdocs and institutions are not local but are national issues. I applied...many changes to the  Board of Directors and the Executive Director changed twice. I had to learn how to ...to the Board of Directors and liaise with the Executive Director. As members changed, I had to learn how...

... sticking pipettes into solution bottles directly, rather decant the solutions into disposable sterile... up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. Twice...that you also have the power to take the added precautions necessary to keep your cells from becoming contaminated...things more manageable, but it also keeps stock solutions from getting contaminated. Make sure cells don't...So if you, for example, split cells on a Monday diluting them 1:10, you should be able to split on the ...

... positives [2, 4]. Solutions to the polymer problem There are some simple solutions available for researchers...GFP to study the protein's behavior in cells, in solution or something in between, you are using a tag with...ruin your day (and experiment) are listed below. Solutions to avoid these all too common issues follow. ...with all of these problems. The simplest and best solution is to use truly monomeric FPs. EGFP and other ...they are designated mEGFP, mVenus, etc. A word of caution is needed because FPs not derived from GFP, i.e...depend on protein concentrations. An imperfect solution is to restrict imaging analysis to cells expressing...

...commonly used for elution, it can give more variable absorbance readings. A Tris-buffered solution tends to produce...readings to average out noisy measurements. Be cautious when interpreting purity ratios, as they can vary...vary significantly at low concentrations.  The elution buffer you use will also affect how accurate your...downstream applications and that you use the same solution for blanking your machine!  Finally, many NanoDrop...systems are no substitute for sound techniques, routine machine maintenance, and critical interpretation...pipette carefully? Remember to discard flow-through solutions and use clean tubes? How about spin times, were...

...which can reduce the potential image resolution of super-resolution fluorescence microscopy. Their size...about 2 nm, which can lead to higher resolution for super-resolution microscopy. Nanobodies can be labeled...animal sacrifice Poor tissue penetration Less resolution due to larger label displacement Require separate...penetration Can be labeled with multiple dyes Greater resolution due to lower label displacement 2° incubation... Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein...

...Blainey, P.C. (2024). CROPseq-multi: a versatile solution for multiplexed perturbation and decoding in pooled...valuable tool for microglia research and broader therapeutic applications.  Find microglia-targeting AAV plasmids...https://doi.org/10.1101/2023.12.12.571321.  Revolutionizing Gene Therapy: The REVeRT Dual AAV Vector System...has developed REVeRT, for gene delivery using reconstitution via mRNA trans-splicing. By splitting a large...Illustration of split fluorophore assay to test reconstitution via mRNA trans-splicing. A cis-splicing vector... The authors used REVeRT to functionally reconstituting coding sequences for a range of applications...mechanisms of delivery and the potential for therapeutic success. Find REVeRT plasmids here! Riedmayr...

...vector solution, and generally these methods measure different characteristics of the solution. These ...can also be altered by freeze-thaw of the viral solution (Lock et al., 2010). Infectious titer: the ...quantified by electron microscopy of a viral vector solution. Since this technique is intensive and requires...requires an electron microscope, it is not routinely performed on all new viral vector preparations. Addgene...facilities report the physical titer of a viral solution (Figure 2). Because physical titers are used for...starting template is the purified viral vector solution (which contains viral capsid proteins) as opposed...

...the complex interactions between cancer cells, therapeutic drugs, and the rest of the body than studies ...or cell culture. For example, Scott Lowe’s lab utilized mice to study how different mutations in leukemia...important human disease model with potential for therapeutic drug discovery as it is estimated that 75% of...they can also be used to easily screen multiple therapeutic compounds in one experiment. Zebrafish (Danio...to name a few. For example, Andrew Leifer’s lab utilized the worm to study calcium signaling in the entire...al. "Whole-brain calcium imaging with cellular resolution in freely behaving Caenorhabditis elegans." Proceedings...Drosophila melanogaster and the role of the fly in therapeutic drug discovery." Pharmacological reviews 63.2...

...Structural problems in academic research and potential solutions During the first couple of sessions of the symposium...his or her chosen career path,” but individual institutions can have many different names for the positions...From Panel Session 2: Defining the Postdoc: An Institutional Perspective. This lack of complete information...workforce, and we are beginning to identify potential solutions. Keynote speaker Paula Stephan did a fantastic... articulating many of these problems and some solutions. The structural problems seem to boil down to ...here). Importantly, the panel offered some great solutions to these problems and some that stuck out to me...those at the FOR symposium and will enact their solutions as soon as they become actionable. Additional...

...cytosine-to-thymine substitutions in target sequences (Banno et al., 2018). These substitutions were limited ...the utility of cytidine and adenine base editors that enable C-to-T and A-to-G base substitutions respectively... is done with recombineering, a technique that utilizes phage recombination machinery to promote homologous...especially for larger modifications. What’s the solution to this inefficiency? Use CRISPR to make it a ...introduce point mutations in P. aeruginosa, Pseudomonas putida, Pseudomonas fluorescens, and Pseudomonas syringae...respectively. Substitutions seem most efficient when using a nickase version of Cas9, but were also successful...

...dramatically reduce CRISPR efficiency, and DNA substitutions don’t fare much better: they largely abolished...recognizes the 2’-OH of RNA in this region, and DNA substitutions dramatically reduced cleavage (Kim et al., 2020...scaffold sequences have different tolerances for substitution of DNA bases. Created with BioRender.com. ...matches, the complex is stable enough that the DNA substitution doesn’t disrupt it. Combined with the reduced...access these kinds of tools. Location of DNA substitution Similar on-target efficacy Reduced off-target...Different targets benefit most from different DNA substitutions, although there are commonalities that may suggest...

...common solutions to avoid background staining when doing species on species IHC. (A) One solution is to...fluorophore) directly attached to it. (B) Another solution is to combine your primary and secondary antibodies...aren’t free to bind endogenous IgGs. (C) A third solution is to use Fab fragments against the sample species...still see non-specific staining! Hopefully, these solutions will help get your experiments back on track.  ... or immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques...

...The most commonly used yeast by biotech and pharmaceutical companies is K. phaffii (formerly known as ...the system. Since then, multiple biotech and pharmaceutical companies (along with some research laboratories...It’s also a methylotropic yeast, meaning it can utilize methanol as its sole source of carbon and energy...distantly related to S. pombe and S. cerevisiae; evolutionarily it evolved much slower and bears more similarity...the expression of recombinant proteins by the pharmaceutical and biotech industries as a protein expression...greatest hurdle for genome engineering. A unique evolutionary feature of K. phaffii, relative to S. cerevisiae...

...display-based directed evolution to create APEX2. They have now applied the same directed evolution technique to...compartments is via proteomics--the study of the distribution and interactions of proteins within these structures...small-molecule substrate to map the spatiotemporal distribution and interaction networks of specific proteins...paper, they describe TurboID and miniTurbo, which utilize non-toxic biotin, but label nearly as quickly as...escape detection. The Alexander Green lab found a solution: a low-cost, cell-free detection assay that can...

...chemiluminescence, precipitate the protein out of solution, or isolate cells expressing this protein. Read... The hybridomas are then subcloned by limiting dilution, which produces an immortalized, clonal population... curve. This curve is generated using a serial dilution of a known amount of protein that is also detected...the protein of interest and pull it out of the solution. Once the unbound protein is washed away, the ...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...investigating signaling pathways or developing therapeutics. Experimentally, antibodies conjugated to fluorescent...

... super-resolution or single-molecule techniques.   Figure 2: Two-color super-resolution imaging...which reacts with O2-benzylcytosine(BC) ligands (Gautier et al., 2008). Although based on the same original...multiple proteins but label each separately before reconstituting, choosing your colors depending on the needs... want to use far-red rhodamine dyes for super-resolution imaging, HaloTag gives better performance than... (2019). Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and...), 547–556. https://doi.org/10.1021/cb900062k  Gautier, A., Juillerat, A., Heinis, C., Corrêa, I. R., ...

... that present as global developmental delays or autism spectrum disorder. It is estimated that CTD is ...could greatly enhance our ability towards CTD therapeutic development,” says Iyer. Since the RESOLUTE plasmid...understand so that we could possibly develop pharmaceutical interventions that would inhibit IL-36 release...says Mellett. This would help the lab identify therapeutic targets in the future. Congratulations Mark Mellett...of developmental delay, and is characterized by autism-like symptoms, seizures, and muscle weakness. So...To study cellular transduction events, we will utilize BRET-, FRET-, and intensity-based signaling biosensors...

... gene expression. On the other hand, using a constitutive promoter in the same background resulted in ...over-represented compared to those further away. While constitutive promoters drove maximal gene expression near...during shifts in growth rate as compared to a constitutive promoter. To simulate the increase in plasmid...When they increased concentrations of IPTG, a constitutive promoter upstream of GFP increased expression...stabilized promoter while gene expression from the constitutive promoter did not. Similar levels of expression... al. placed the genes under the control of a constitutive promoter and then moved them into the genome...

...human conditions, opening up new avenues for therapeutics that treat disease at the molecular level. For...patients have achieved persistent immunological reconstitution with a normally functioning T cell repertoire...design to clinical trials involve identifying a therapeutic gene, finding a means to deliver it and identifying...issue by developing new capsids through direct evolution, capsid shuffling, and peptide displays. These... and other ongoing clinical trials support the utility of AAV as a broadly applicable gene delivery tool...understanding of disease pathogenesis will enable robust therapeutic outcomes. In contrast to other gene delivery ...

...can be removed either by filtration of the virus solution (through a 0.45 µm or 0.22 µm filter) or by a ...sterile PBS.  Alternatively, to concentrate a virus solution without ultracentrifugation, you can reduce the...the virus is released, a more concentrated virus solution can be obtained. 4. Enhancing virus-cell contact... in single-use aliquots. Never use a polybrene solution that has been thawed more than once. Fibronectin... packaging cells. This check can be performed routinely and without disrupting the flow of your experiment... of the viral vector using the harvested viral solution as a PCR template. Regardless of the type of virus...