... is our own immune system, which can recognize antigens on cancerous cells and eliminate them from the...ensues, traditional chemotherapies have taken advantage of the universally-dividing behavior of cancer...approaching the Ras problem by exploiting the different mutant versions of Ras that are most prevalent in human...fluorescent protein fusion studies and the generation of mutant cell lines. Further, for each plasmid in the kit...making the results of any such study as clinically relevant as possible. Connecting the research community...community, will soon expand to include several other important pathways. We invite feedback and contributions...of signaling pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014...

...of detection; they are useful almost anytime you want to express multiple genes in the same cell. Although...expressed from the same cassette is sometimes advantageous, particularly when only a portion of the plasmid...expression levels between two or more genes is important. How do multicistronic vectors work? Scientists... mRNA. However, some bicistronic vectors take advantage of an element called an Internal Ribosome Entry...bicistronic vectors; however, they do have some disadvantages. These elements are quite large (500-600 bp)...vector.3  2A Peptides To overcome some of the disadvantages of the IRES element, scientists have adapted...PubMed. 5. Scalable signaling mediated by T cell antigen receptor-CD3 ITAMs ensures effective negative selection...

...derived. For a breakdown of GFP, it's variants, and their relevant mutations, check out Marcy's post from...FPs just keeps getting larger with hundreds of variants available. Read on to familiarize yourself with...emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP (CFP), and yellow FP (YFP...calculated as the product of extinction coefficient and quantum yield of the protein, divided by 1000. In many ...temperature). pH Stability: This parameter is important if you are planning to express the FP in acidic...with very comprehensive data sheets on many FP variants Plasmids 101: Green Fluorescent Protein (GFP) ...

...it is limited to only 20 bases, or why it’s so important to choose a target sequence with a nearby protospacer-adjacent...unpaired strand in the center of the protein. The repeat:anti-repeat duplex pokes out from the REC lobe and...blue.   The REC lobe binds tightly to the repeat:anti-repeat duplex of the guide RNA, confirming its..., but they improve cutting efficiency and are important for robust activity in cells (Jiang & Doudna, ... loop to produce SuperFi-Cas9, a high-fidelity variant that still retains high cleavage efficiency (Bravo...correct positioning of the non-target DNA strand is important for stabilizing the active conformation of the... and different techniques all bring their own advantages—and blind spots—to these questions. Each new ...

...book left them wanting to learn more. There are individual paragraphs that may have warranted their own chapters...politicization of science (from climate change deniers to anti-vaxxers), and the very real dangers of quackery,...self-promotion, and personality is playing an increasingly important role in science. The science produced by the quieter...etc.) with that produced by the confident and flamboyant Watsons.” In Part Four, “New Science,” Zimmer...posits the way science is being done has changed significantly over the last decade. He describes the detection... chapters, and certainly chapters that could warrant their own books. Multiple books have been written...

...create specific edits in DNA, there has also been significant work on constructing Cas9 transcriptional activators...upregulation of essentially any gene by fusing mutants of Cas9 deficient in DNA cutting activity to a ... working as expected. Finally, for each gene you want to activate, you should also be ready to test three...Thus, in cases where users have a single gene they want to activate, we would recommend using a cDNA overexpression... the promoter of another gene, thereby driving aberrant transcription. That being said, we generally pick..., Picco G, Gonçalves E, Beaver CM, Migliardi G, Santos R, Rao Y, Sassi F, Pinnelli M, Ansari R, Harper...Addgene Blog Learn how to Choose the Best Cas9 Variant for Your Next Experiment Find out More about Genome-wide...

...future.  Many thanks to our guest blogger Samantha Young. Samantha Young is a Medical Writer with a PhD in... This post was contributed by guest blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has...al., 2013, Cho et al., 2014). There are several variants of the CRISPR/Cas9 system floating around, and...and many pre-designed plasmids containing these variants ready for purchase. But what is the easiest and...). In this case you should be able to find your mutant from screening as little as 8-10 pups. The level...PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice by pronuclear injection of circular plasmid...

...compelling of these were those from researchers who wanted more time to mine their own data, with some smaller...work this may entail.  So if access to data is important, what data should be included? We should recognize... storing data in a sharable form is indeed a significant effort. Our standards of data recording are highly...encouraging authors to integrate datasets with their resultant figures. Instead of providing a “dead end” figure...touch interfaces where the viewer is more likely to want to manipulate how they view the information. We ...importance of data sharing The reason this topic is important to my lab is three-fold. Firstly, we rely on many...experiments and understand their data. Likewise, we want to contribute to helping others understand ours....

... the brains of living mice both quickly and significantly. Genes under LITE control showed a rise in transcription...epigenetics with the addition of histone modifiers. “We want to allow people to prove the causal role of specific... Möglich and Peter Hegemann described several advantages of the new system: "First, the light-responsive... methyltetrahydrofolate, which are universally abundant. Second, induction of transcription occurs within...in C. elegans and in Plasmodium. For those who do want to apply LITEs for the transformation of neurons...tip: save yourself some time by making an AAV1 supernatant. (The details can be found here.) So tell us,...

...methylase mutation (GATC) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes... mutation (CCWGG) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F-mcrA Δ(mrr-hsdRMS-mcrBC...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F- gyrA462 endA1 glnV44 Δ...maintenance of a plasmids with R6Kγ origin; contains a mutant allele of the pir gene that maintains the plasmids...screening and routine cloning, nalidixic acid resistant. endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44...plasmids, ligated DNA, and libraries, nalidixic acid resistant.  endA1 glnV44 recA1 thi-1 gyrA96 relA1 lac Hte...

...relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said...comprehensive understanding of how cancers become resistant, an emerging theme is that they do so by activating...signaling pathways involved in drug resistance Wood wanted to find out which particular pathways drive resistance...pathways, they identified a set of one to three mutant complementary DNAs (cDNAs) representing core nodes...of - and that’s thrilling to us,” Wood says. “We want to make these available as widely as possible.” ...of signaling pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014...

...annotate genetic perturbations that may play significant roles in cancer pathogenesis. Their work culminated... any mutations that that confer proliferation advantages will be revealed in the absence of growth factor...mutations were found to be “neutral,” having no significant effect on cell viability compared to a wild-type...Approximately ~22% of the mutations, however, significantly increased cell viability, and these “activators...passenger mutations from the “active” drivers is important for the development of targeted therapeutic interventions...dozens of wild-type genes and their associated mutants in other model systems. It is possible that a given...

...using Twitter to find phages to treat an antibiotic resistant bacterial infection, she shared that tweet...for phage may not be needed if other antibiotics work. “We don’t want to cry wolf to the phage community...woman with CF whose infection has failed all #antibiotics. We need lytic non-lysogenic phage URGENTLY to... a repository of phage data and information. “We want to be the go-to place for phage information because...

...(IPI), in which Addgene will distribute recombinant antibodies (rAbs) developed at IPI. The first IPI ...excited to launch our Antibody Data Hub, an open-access repository for antibody validation data. We’ll...world. That included 2008 pooled libraries, 290 antibodies, 18,359 viral items, and 166,247 plasmids. We... our distributor partners, with the launch of antibodies in China and expansion of viral services available...

...scale: Look at the mEmerald variants first, followed by the color variants. The mEmerald plasmids offer...Davidson lab prepares to shutter its doors. “We wanted to give all of these vectors to the scientific ... FSU. Mike is really the one who had been very adamant that we distribute to as many researchers as we...reagents. "We are proud to be able to bring such an important collection to the scientific community," says ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

... have pushed the boundaries in neuroscience by granting targeted control over neuronal activity. While..., both techniques offer researchers different advantages and limitations.  The four main factors in which...studies, however, require a permanent intracranial implant to be surgically installed to allow for light delivery... delivery. Chemogenetics poses the advantage of being less invasive and more flexible as it relies on ...Invasiveness Mice require permanent intracranial implant Noninvasive Controlling Stimulation ExogenousEasily...controlled EndogenousNot easily controlled Unique advantages Mapping the brainDissecting circuits Studying...

...direct, specific and sensitive detection of the plant hormones strigolactones By Fabienne Furt Current... detect strigolactones (SLs) are costly, non quantitative or require heavy optimization. Claudia Vickers...fluorescence intensity, it provides a direct and quantitative detection of SLs. The authors also showed that... known to trigger seed germination of parasitic plants, this approach was also used to build and validate...Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and reduced...

... our ReFigures on the therapeutic use of anti-CD47 antibodies in immuno-oncology. Funding agencies such... Cataloging a reagent’s performance as in this antibody ReFigure Analyzing replications as in this ReFigure... ReFigure to coherently save analyses with the relevant figures from a literature review. No more hunting... and good for science ReFigures are published instantly and are editable by the author at any time. Insights...preprints, data, software, micropublications) in grant proposals. ReFigures are one such research product...

...ve realized my figures are just as important, if not more important, than my writing. Initially I didn...design software, like Adobe Illustrator, and I didn’t want to pay for these programs either. But with a little...presentations, manuscripts, or social media. If you want to draw your own figures Google Drawings Google ...through your university or your public library. If you want to use pre-drawn images in your figures Biorender...New icons can be requested, although there’s no guarantee Biorender will create it. Additionally, you can...

...ever been the target of an ‘-omics’ study or is resistant to specific compounds, links to publication references...cross-references with cell line catalogs and collections. Importantly, Cellosaurus also identifies problematic cell... the most common contaminator line.  For those wanting to access Cellosaurus programatically, their API... helpful information on the known features and relevant literature, or to decide which line is best for...scientists all over globe, with some surprising, and important, findings being shared. For instance, recently...around! If you’re excited about this resource and want to learn more, check out the Cellosaurus FAQ! References...