...incredibly useful transferable skill. Being able to stand up in front of an audience and convey your ideas...you need it, rather than trying to push through a state of panic. Now, if I feel myself getting too nervous...present a paper to our classmates so they could understand it. When my mind went blank, my options were ...sense. As tempting as option two was in my panicked state, only option one would let me accomplish my goal... me.) They happen to everyone! Everybody makes mistakes, encounters technical difficulties, and runs into...you or not going perfectly, you truly are a rock star.  The best thing about public speaking as a scientist...

...and SMASh degron tagging vector! Destabilizing domains Destabilizing domains (DD) are akin to degron ...into the system, it will bind and stabilize the DD, resulting in sustained protein levels. Looking for a ...turning on expression of a new protein is a cinch. Stably introducing the coding region of a gene (cDNA) ...mechanism. When a protein is tagged with a DD, it is unstable and quickly degraded. However, if the relevant...

...Schiebinger, PhD, Hinds Professor of History of Science, Stanford University. Sex and gender are critical components...Commission, the US National Science Foundation, and Stanford University—that harnesses the creative power of...Gendered Innovations. This project: 1) develops state-of-the-art methods of sex and gender analysis and...sexes come from, one clue or potential place to start research is by considering sex disparities in disease...important point, however, is that cost at this early stage of research is minimal compared to that of human...trials. And, as Janine Clayton points out, late-stage clinical studies that draw from a body of research...

...each ligation reaction uses a pool of gRNAs as starting material, the ligation products are a diverse ... al., 2014) and microRNAs (Wong et al., 2015). Starting your CombiGEM-CRISPR experiments The first thing...efforts made by various research teams to build and constantly improve gRNA libraries, excellent resources with... Wong et al 2015. Alan Wong is currently an Assistant Professor at the School of Biomedical Sciences... applying new technologies to interrogate and understand complex biological systems. He can be contacted...Timothy K. Lu. "Enhanced killing of antibiotic-resistant bacteria enabled by massively parallel combinatorial...

...to discuss their visions for a successful and sustainable biomedical research sector at our first Future...at this event added to a growing dialogue on the state of a system that, despite all of its successes, ...years of earning their PhDs (3), showing that the ‘standard’ pathway into academia may in fact be just another...Andrew Young School of Policy Studies at Georgia State University, is an expert on the academic labour ...postdocs. Among other involvements, he helped to establish grants through the Sloan foundation for postdoc-related...uses synthetic biology to create tools to better understand human disease.  Outside the lab, he is interested...

...require protein interactions to remain stable (or to be stabilized) during cell lysis and lysate fractionation...identification techniques rely on protein interactors to stay in close contact with the bait protein throughout...can artificially select against PPIs that aren't stable outside the native cell context. The Split-BioID...expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown in biotin-containing...characterization of these candidates is critical to understanding how they interact with the bait proteins. Another...

... addition to sequencing plasmids, we have also started using NGS to check the quality of other services...quality data! Addgene's new in-house NGS process To start, the process is only made possible by the use of...quickly create Illumina sequencing libraries. From start to finish the library preparation process only takes... individually assembles the reads into a single FASTA sequence that our QC scientists can easily analyze...performed, optimized as needed, and cleaned up using a standard PCR cleanup kit to remove any template DNA and...as part of our OpenBio Repository sometime soon. Stay posted!     References Guerin K, Rego M, Bourges...

...-fold more photostable and as bright to mVenus, the parent YFP, and is the most photostable YFP to date... were exposed to blue light. Cre-Vvd showed substantially improved activity as compared to other recombinase...addition, Casɸ has a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where....ncbi.nlm.nih.gov/32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin ...contribute to experiments that require high photostability, such as time-lapse imaging or with high light...

... biomedical innovations promise an improved understanding of how our bodies work, new treatments for debilitating...School, has a mission of raising awareness and starting conversations about both the benefits as well ...between these communities and the biomedical establishment. This, along with concerns about the use of ...year, I’ve since returned to pgEd as a full-time staff member for the past three years. pgEd is a small...diverse backgrounds and disciplines, and everyone on staff takes on a variety of projects. With our focus on...submitting op-eds to your local newspapers, or starting your own blog. Even if you do not end up pursuing...

...laboratory in a cell-free and biosafe format that was stable for up to a year at room temperature (Pardee et...gene networks for the detection of antibiotic resistance and strain-specific diagnostics for the Ebola...Pardee and Alexander Green! Keith Pardee is an Assistant professor in the Leslie Dan Faculty of Pharmacy...the environment.     Alexander A. Green is an Assistant professor at the Biodesign Center for Molecular...and the School of Molecular Sciences at Arizona State University. His group carries out research in synthetic...self-assembly for applications in health, energy, and sustainability.   References 1. Pardee, Keith, et al. "Rapid...

... Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires...nt upstream of the PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18... 18-23 bases away from the PAM. Cpf1’s staggered cleavage pattern opens up the possibility of directional...efficiency of these two processes will help us better understand where Cpf1 fits into the genome editing landscape...Addgene. 4. Makarova KS, Wolf YI, Alkhnbashi OS, Costa F, Shah SA, Saunders SJ, Barrangou R, Brouns SJ,...

...101 eBook to get all the background you need to start working with plasmids! What are termination and ...structure of the hairpin-DNA complex is thought to destabilize the transcription complex, initiating cleavage...addition of the poly(A) tail is important for stability of the mRNA, protection from degradation, and ...vectors often employ other non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) ...Maitland NJ. An Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered...Addgene.org Browse our Plasmid Reference Pages Just Starting Work in a New Field? Check out Our Plasmid Collections...

... mostly learned through experience and from my mistakes. Here I’ll share what I wish I knew when I first...He left after a semester while the other student stayed until he graduated. Years later I attended a formal...often and clearly As a manager you need to clearly state expectations and goals. Yours and your team’s success... can be hard - but it is the best way to avoid mistakes due to miscommunication and to detect trouble....grateful for the feedback, but at least you can start making changes for the best. Positive feedback is...encourage them to speak their minds. They will make mistakes and you will take responsibility (within reason...

...outside a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing cells for...identify new target genes for drug discovery or to understand the intricacies of molecular pathways. These ...available at Addgene. The large packaging capacity and stable expression of lentiviral vectors make them an attractive...first two steps, as no mice are involved. Ex vivo starts with transducing cells with the CRISPR library ...options involve generating strains of mice that stably express Cas effectors in specific tissues, eliminating...

...the mice, and restraint method. Many different substances can be injected, including cells, drugs, DNA,.... IV injections are used for a broad range of substances, as it circulate what is injected throughout ...technique. Intrathecal injections are used when the substance being administered must be delivered to the central...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...invasive, and only requires a pipette. Since the substances are administered through the nose, they also ... & Vasbinder, M. A. (2011). Administration of substances to laboratory animals: routes of administration...

...or looking to improve your current methods, understanding the differences can help you make an informed...relatively quick. After transfection, HEK-293T cells start producing virus within 24 to 48 hours. Harvesting...specific application. With mammalian systems, it is standard to do only one passage (transfection). This differs...can even produce a P3 virus! Time Sf9 cells will start producing virus within a day or two, but unlike ...depending on the protocol used. Plaque assays are standard but can take seven to 10 days. Commercially available...Tompkins, G. J., & McCawley, P. (1977). The establishment of two cell lines from the insectspodoptera ...

...antigenic drift.    Starr et al Cell. 2020. https://doi.org/10.1016/j.cell.2020.08.012  Starr et al Science....RBD DMS Library generation and sequencing. From Starr et al Cell., 2020. (B) Mapping antibody-escape mutations... mutations with the yeast display library. From Starr et al Science., 2021. SARS-CoV-2 infects host...cell, improving biosynthetic pathway yields by stabilizing rate-limiting enzymes. In this method, the major...gene. SlugCas9 is a compact Cas9 ortholog from Staphylococcus lugdunensis that recognizes a NNGG PAM and ...

...provide a large transgene packaging capacity. For instance, with a packaging capacity of about 6 kb, the ...studies, the strategy employed was to deliver a standard AdV vector to the targeted organs or tissues with...several biological systems. It has been used for instance in inherited disorders such as B-thalassemia, ...labs wishing to create this kind of animal models. Start using adenoviral vectors with your CRISPR/Cas9 research... (2016) Viral vectors for gene therapy: Current state and clinical perspectives. Biochemistry Moscow 81...Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering...

...2017 and updated Jul 2, 2020. Promoters may be the star of gene regulation, but enhancers and chromatin ...original CAPTURE method requires the creation of a stable cell line that co-expresses three components: ...ligation creates chimeric circles of DNA that are distant linearly but close together in space. The chromatin...CAPTURE 2.0 components eliminates the need to create stable cell lines and allows CAPTURE 2.0 to be used with...BirA Endogenous biotin ligases Expression system Stable cell line expressing 1) FLAG-biotin-tagged dCas9...accessibility, and epigenetic modifications, we can start to build a timeline of events that surround changes...

...Geobacillus stearothermophilus is more stable in human plasma than the standard spCas9 (Harrington et al. 2017...transfection (B in the figure above), are in the early stages of development. David Liu’s Lab has demonstrated...experiments may not be suitable for experiments where stable Cas9 expression is required, such as when a Cas9... little is known about the immunogenicity and stability of the Cas9 protein in the host. Indeed, research...PMC5253684. 5. Harrington, Lucas B., et al. "A thermostable Cas9 with increased lifetime in human plasma...