...Despite these grim statistics, new strategies for attacking cancer are yielding promising results in the laboratory...and balances: protecting against cancerous cells Luckily, there are several safety mechanisms in place to...universally-dividing behavior of cancer cells, attacking the cellular machinery necessary for division....in the field of cancer research has focused on picking apart the complex cellular pathways involved in...

...consider a dual-sgRNA system. This system uses a “nicking” Cas9, in which one of Cas9’s two critical enzymatic...deleting the catalytic residue of a protein or knocking out a very small peptide. In these cases, you ...sites was sufficient to repress transcription by blocking its initiation. In mammalian cells, robust repression...interest. dCas9-based repressors are useful when knocking out a particular gene isn’t feasible. For example...

...the intensity of fluorescence signal and enables tracking of single molecules within living cells without...roles in many cellular functions, from endosomal trafficking to signaling pathways to the cell cycle. Good...used with your own reporters to decipher their trafficking routes. This collection is now available at Addgene...traditional cloning strategies may seem preposterous. Luckily, the Pierre Neveu laboratory designed a new iterative...and an endoplasmic reticulum export motif and trafficking sequence for improved targeting to the plasma...studying membrane dynamics, for monitoring cargo trafficking, and for discovering organelle contact sites..... This appears to contribute to more efficient blocking of read-through transcription for pXPG. The other...

...mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 12–...

...self-excising cassette (Dickinson et al., 2015) made this step as simple as heat-shocking to induce the expression...2013), or rescue of mutant phenotypes (unc-119) (Dickinson et al., 2013). These approaches allowed effective...e193–e193 . https://doi.org/10.1093/nar/gkt805 Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein...

...localization, turnover, and trafficking, as well as organelle trafficking and dynamics (fission, fusion...for whole cell imaging and studying organelle trafficking. Monomeric PA-FPs are usually preferred for studying...Photoactivatable GFP tagging cassettes for protein‐tracking studies in the budding yeast Saccharomyces cerevisiae...

...dilutions. Pro-Tip Make sure that the qPCR is valid by checking to the following: Standard curve: R 2 (coefficient...

...immunohistochemistry images that are crisp, bright, and lacking in non-specific staining can be a challenge in ... mixing your primary and secondary antibodies, blocking excess secondary with serum from the same species...0022155413511620  Lu QL, Partridge TA (1998). A New Blocking Method for Application of Murine Monoclonal Antibody...

...contrast” variant provides improved detection for tracking large populations of neurons in wide-field imaging...fusion fluorescent proteins can affect DREADD trafficking and localization. The Sternson lab recently deposited...-CamKII-ArchT-GFP (PV2527) 105679  AAV5 ppAAV-CKIIa-stGtACR1-FusionRed New tetracycline transactivators...

...throw a therapeutic monkeywrench into the virus’ hijacking of human cells. Even better, more than half the...the Krogan lab's Addgene page to get notified!) Attacking coronavirus, one FedEx package at a time Since...headache receiving constant email requests and tracking their fulfillment manually in a shared spreadsheet...

...components: the error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis with error...error-prone polymerase (PolI) that is fused to a nicking Cas9. A guide directs Cas9 to a site of interest...changes promote nCas9’s dissociation from DNA after nicking and yielded an enhance nCas9 (enCas9) EvolvR with...

...bind to amine groups present in some conjugates, blocking the conjugation reaction. Many commercial antibodies...for your ELISA and Western, I’d highly recommend sticking with what works.  To balance this out, however...in your machine before you make your purchase. Picking a single fluorophore for an experiment is easy,...

...definitely recommend checking out GCE4All’s website and resources, and then checking out all of the plasmids...focus on development, along with the inherent trickiness of using and optimizing GCE, led to a large toolbox...

...genome randomly, while others use attP sequence docking sites so that you can control exactly in which ...at different loci. Fortunately, there are many docking sites available for integration. 2. How soon would...would know beforehand on which chromosome your docking site is located). This amounts to a small difference...

... it’s monumentally difficult without software. Tracking lab procedures also gives you access to hard data...repeated. We also now have a good interface to begin tracking analytics data for lab procedures. With this data...are distinct from their contents. You should be tracking both, you should easily be able to move contents...

...localization in cells, or even single-particle tracking of the bound complex. There are so many FPs available...TagBiFC technique allows long-term single-molecule tracking of protein-protein interactions in living cells... can even be bright enough for single-molecule tracking in live cells.  Controls  When designing experiments...

...Additional opportunities: Tracking intracellular pathogens In addition to tracking the location of proteins...University of Queensland) to observe membrane trafficking events in vivo. Locate your protein and see who...

...Miyatsuka et al.) Distinguish between models for trafficking to the lysosome by fusing Medium-FT to LAMP-2A...structure and a particular biological event (trafficking to a subcellular location, start of gene expression...change color from blue to red report on cellular trafficking." Nature chemical biology 5.2 (2009): 118-126...

...programs is almost as daunting as the program itself. Picking a mentor comes with many challenges, and the flood... to potential mentors?” “Where do I start with picking universities?” and “How does COVID-19 impact all...into what matters to you. It can also be nerve wracking to ask questions in front of other prospective...

...in “crispr” and see what comes up. Before even clicking the ‘Search’ button a new content-aware suggestion... your lab. Instead of starting a new search or clicking through to a different part of the website, you...functionally (e.g. Base Editing) and by target organism. Clicking through to RNA Editing brings us to a page with...