...molecule integrin inhibitors and natural ligands, such as fibronectin, to block receptor function, e.g.,...technologies and laboratory techniques related to antibody discovery. You can find cell surface receptor biology...Integrins are essential cell surface receptors involved in complex signaling pathways and the linkage...Despite their ubiquity and significance, these receptors have proved challenging to study. Cells often ...integrin antibodies available through Addgene’s repository. The collection includes 26 unique antibodies...from its closed to its open state, activating the receptor. Ligands can bind to integrins in their closed...subunit. Using these affinity reagents, the investigators inhibited other RGD-binding integrins, individually...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...activation, using expression of differentiation factors as the screen readout. This retroviral mouse CRISPR...library can be used for screening of cells that are refractory to lentiviral transduction, now enabling forward...lipoproteins, shuttles lipoproteins through the circulatory systems, but can also lead to the formation of...as both a powerful tool for generating complex regulatory systems and further the study of natural σ54-... PMID: 31451697. Genetically encoded voltage indicators for use in worms Article contributed by Brook...Brook Pyhtila Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when neurons...

...the abundance of a protein variant library is monitored by fusing the variants to eGFP followed by an ... is an enzyme critical for activating clotting factors and is the target of the anticoagulant drug warfarin...serotonin sensor The goal of Elizabeth Unger’s postdoctoral work in Lin Tian’s lab at UC Davis is to generate...serotonin is a well-known but poorly studied neuromodulator, and evidence suggests that it is incredibly...sensor). In preliminary studies with the lab’s collaborators, they were able to use the serotonin sensor ...activity. Unger also plans to use the serotonin receptor to study how single nucleotide polymorphisms present...and dose response. Selective serotonin reuptake inhibitors are frequently used to treat depression and anxiety...

...upon a moratorium until another meeting, three branches – a private funder, government regulators, and ...there was a generally agreed-upon international moratorium on utilizing CRISPR/Cas9 in humans. This topic...mtDNA transfer options being investigated; A UK regulatory body tasked two separate non-governmental groups...technology to research centers engaged in long-term, mandatory follow-ups and observational studies of the technique...whether or not scientists can recognize so-called bad actors among them and “self-police” or whether the media...to argue otherwise). To be sure, throughout the history of medical research, there have always been people...stop punting decisions on scientific, ethical, regulatory, and general public consensus to the next meeting...

... fields, including synthetic biology. Addgene depositors Christopher Voigt and Jeffrey Tabor have been...and inactivate biological processes. Chemical effectors are suboptimal for a number of reasons, including...cross-reactivity with other pathways. Since these effectors are diffusible, they’re also not suitable for ...histidine kinase that phosphorylates a response regulator (RR). TCS-induced responses are varied; one outcome...conditions, the chromophore activates the response regulator via phosphorylation, promoting transcription of...spread across three plasmids, with the response regulator encoded in the genome. To make the system less...The utility of these TCSs was limited by a few factors. These bulky systems were spread over a number ...

...effect of mutations of these regulatory elements on gene expression. The regulatory element (wild-type or mutant... from live cells. Monitor miRNA knockdown Expression of luciferase can be monitored as a downstream measurement...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection...Kinetics of Photon Generation Slower Faster Cofactors/Substrates Required Not Required Signal Strength...Organism Peak emission (nm) Size (kDa) Substrate Cofactors Secreted Luminous shrimp(Oplophorus gracilirostris... most often used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated...can then be used to quantify the effect of the regulatory element. Depending on the specific luciferase...

... important, factors to consider when seeking your dream lab. The principal investigator's point of view... hires are simpler because of their documented history in science. Potential graduate students typically...goals and chat about awesome science. Important factors in the search for a lab Studies show that job satisfaction...both work style and development needs. These two factors will provide a solid foundation on which to thrive... this should (in most cases) not be the driving factor. The following questions can serve as a guide for...you take the position. Am I choosing the right mentor? Make sure your advisors will be good role models...more on this topic see our blog "Choosing a Good Mentor for Scientists". This post also includes a useful...

...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the...functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene of interest is inserted into the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...plasmid does not contain the insert. If you’re using vector-specific primers, look for size differences. You...

... your gene of interest in a plasmid and get the vector incorporated into the germline. To begin this process...knockdown. Addgene also has a number of insect vectors to consider. An easy way to see what is available...Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important...important points to consider when choosing the vector you’d like to use to generate your new fly line:... gene to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others...would you like to select for your flies? Sure, the vector will have an antibiotic resistance gene to select...transgene. One such marker is the white gene, which restores eye color to fly lines where the endogenous white...

...project management. Mentorship is another invaluable resource: connecting with mentors who have made similar...management often means stepping into unfamiliar territory. Unlike solving technical problems in the lab,...lab, management involves a different skill set: mentoring, setting strategic goals, and fostering team success...for me. I began exploring alternative career trajectories within the industry, but the transition wasn... role is also crucial. I owe quite a bit to my mentors, as they encouraged me to explore small-scale opportunities...company by taking on responsibilities such as mentoring a junior colleague. Guiding an intern, in particular...and leveraging tools like the Myers-Briggs Type Indicator or DISC assessments can help you navigate team...

...feels that storytelling is an impactful tool for science communicators. “The power of a story is to make... to learn things quickly. Story telling. The ability to know what’s a story and how to hook your readers...there’s always plenty of writing: plain-language stories about Fred Hutch research, pieces for donors such...that can be a very painful process. And certain stories can make you feel like ‘I’m a terrible writer. ...through connections and that’s something that good storytellers in science can do.” Susan’s number one piece...social media as a scientist here Learn more about storytelling in science ...

...AMA I am a strong advocate for peer mentoring, but volunteer mentors are only able to go so far with their...coach do that is different from an adviser, boss or mentor? First, professional coaches have experience and...infrastructure are not always as effective in mentoring for this transition. Second, a coach will help...experiences. Good coaches will have loads of success stories and plenty of people to vouch for them. Trust...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...online content for PhDs and postdocs posted by Toronto-based Jennifer Polk of From PhD to Life. She runs...has an outstanding blog and, while her PhD is in History, I think she would be a good match as a coach for...

...scientists who were potential Addgene requestors and depositors. China is in one of the top 10 countries...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to ...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...human tRNA genes and, until now, we've overlooked regulatory fragments of these transcripts. I remember when...was kicked off by Nobel Laureate (and Addgene depositor) Craig Mello. Taking his groundbreaking work in...Medicine presented another unexpected finding in the regulatory RNA field. The Rajewsky lab is studying circular...

...utility of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses... here! Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School ...in immune cells have identified many putative regulators of immunity. However, in vitro screens cannot... of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout ...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...responses to cancer. The screen recovered known regulators, such as Ptpn2, Pdcd1, and Zc3h12a, demonstrating...

...into wide use, there were a variety of ways to monitor cell, organelle, and protein localization. For ...the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers can...Tsuboi et al.), limiting its use. Nonetheless, its creators noticed a few properties that have now become ...benefit of the common FP timers is that you can monitor age by expressing a single FP-timer protein or ... These separate proteins should be useful for monitoring cellular events occurring at varied time scales...fluorescent protein timers FP timers have been used to: Monitor dynamics of vesicle fusion and release from the...vesicle cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...

...Administration of substances to laboratory animals: routes of administration and factors to consider. PubMed, 50...type of injection to use depends on a number of factors. You have to weigh the pros and cons of various...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...common use of intradermal injections is studying inflammatory responses, as this method is useful for analyzing...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...

...bacteria, but have no fear - plasmids from Addgene depositors are making it easier than ever to use CRISPR  ...double stranded breaks (DSB) are lethal. Addgene depositor Luciano Marraffini’s lab took advantage of this...activation of transcription including transcription factors, phage proteins, and RNAP components (Dong et al... al., 2018). They found a transcription factor called SoxS for CRISPRa. Adding complexity, they found ...been observed with Phagemids (Selle et al., 2020, Citorik et al., 2015, Bikard et al., 2014). When the CRISPR... on the presence of specific target sequences (Citorik et al., 2014). These could be genes, specific polymorphisms...refined by the addition of a uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit...

...different Cas12a inhibitors. Some of these anti-Cas12a proteins have broad inhibitory effects on Cas12a...Caixia Gao’s lab has developed a new plant base editor from an evolved tRNA adenosine deaminase that is... is fused to a nicking Cas9. This base editor can be used for A•T to G•C edits in protoplasts or plants... Biology Find plasmids containing these base editors Spatially and temporally regulated AAV-based ...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics and a screening approach...

...zinc finger nucleases to transcription activator-like effector nucleases (TALENs). Our most popular kit... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these two...now have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene...be popular years after they were added to the repository, including beta-catenin reporter M50 Super 8x... GCaMP6 series of genetically encoded calcium indicators were the hot technology of the year in our collection...

...DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering... “Transcriptional activators are easy to expand or combine with different effectors. I’d encourage people...The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn...devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...expression levels reliably increased in vitro by a factor of 10 to 20. A flexible and modular optogenetics...