...manipulating interneurons using a viral approach Genetically modified organisms like transgenic mice are important...a synthetic red fluorescent protein variant they call mScarlet. Bindels et al used a combination of deliberate... cell physiology and molecular processes via genetically encoded proteins. PhoCl is a new member of this... such as an NLS or NES, enables control over localization. Inserting PhoCl between an enzyme and an inhibitor...experiment, check out our recent guest blog series: A Practical Approach to Choosing the B(right)est Fluorescent...investigated an aspect of telomere length regulation called telomere trimming and have proposed a role for ..., Li and colleagues observed endogenous TZAP localization at telomeres and found that TZAP binds directly...

...limitation and allows for the flexible design and small scale production of antibodies. Problems with antibodies...instead of via recombinant DNA, they can’t be genetically tagged and instead are conjugated to reporters...labeled tyramide molecule into a highly reactive radical, which is then covalently linked to residues within...however, compatible with most multi-labeling immunohistochemical protocols where either a directly labeled... required for detection RANbody is detected enzymatically with HRP substrate More sensitive than detecting...RANbodies) as versatile, small, sensitive immunohistochemical reagents. PNAS. PubMed PMID: 29440485. Additional...

...composed of four peptide chains (two identical heavy chains and two identical light chains) each with a constant...antibodies means that it is relatively easy to physically divide up the different regions, both through...joined by a linker peptide, resulting in a product called a single-chain Fragment Variable (scFv). Size: ...samples more efficiently and can also improve localization accuracy when used in super-resolution microscopy...David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody Fragments. Antibodies 8(...

... audience and convey your ideas clearly and strategically is powerful in almost any career. It allows ...my head. What came out, though, was an incredibly calm: “Excuse me, I seem to have lost my place. I need... the open windows in the middle of your talk. Technical difficulties turn your slide deck into a chalk...everyone! Everybody makes mistakes, encounters technical difficulties, and runs into unexpected stumbling..., to a group of peers who are there to provide critical feedback, often with a semi-international audience...your own native language. And, at least in the biomedical field, with very little text on the slides and...

...your knowledge at local schools. Or work as a guide in local museums, zoos or botanical gardens. You can...unusual setting: Discuss your latest research in a local pub at a Pint of Science event. Or get on stage ...to reach young people? Try networking with your local education community. Get involved in education advocacy...communication organisation like Building with Biology. Local environmental groups, charities and NGOs are often...

... have a quiet place to take your video call.  Expect technical difficulties. To try and circumvent this...ago. I just recently started working in the pharmaceuticals industry after earning my Master’s in 2016 ...presentation as an opportunity to demonstrate technical and transferable skills that make you qualified...perfect science talk Be prepared for potential technical difficulties. Have your presentation on a flash...from HR carefully. And even if you don’t have technical challenges, your interviewers might.  You’ll probably...

...Mother Nature’s toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid...carrying genes for bacterial toxins like colicin typically also carry the necessary resistance genes to help...cells survive their own secretions. Pro tip! Historically, colicin-producing plasmids have been referred...which strains of E. coli contain this F plasmid is critical for successful Gateway cloning. In addition, scientists... Vogel, T. M., & Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer in...Characteristics and Potential Applications in Biotechnology. Critical Reviews in Biotechnology, 24(4), 155–208. https...

... be used for measuring protein expression or localization in cells, or even single-particle tracking of...named according to the number of strands in the canonical beta-barrel structure. Most frequently, the proteins...fluorescence complementation analysis under physiological conditions. Shyu YJ, Liu H, Deng X, Hu CD. Biotechniques...fluorescent proteins as tags and biosensors for multiscale imaging. Shcherbakova et al. Nat Commun. 2016...split-FP options. While the BiFC interaction is typically irreversible and requires some time for chromophore...

...very useful in the event that you will not be physically present at your poster at all times, so other...readers can navigate your poster and follow the logical progression of your work, and you can accomplish...offers free poster templates, although most are vertical.  Large central box. Another commonly used and...presentation. Portrait. This style displays a poster vertically rather than horizontally. Instead of displaying.... Shapes. In this type of poster, written and graphical information are inserted into larger circles or... or ovals, that are arranged in some logical pattern across the poster. This style has the advantage of...

...histone-derived peptide) which can localise the construct specifically to the histone protein of interest...becomes. This prevents the cellular machinery from physically interacting with the genes in the compacted region...grandchildren; in this study,epigenetic changes were statistically significantly associated with reduced life-span... Chi-Wang, Cindy Y. Jao, and Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation...methylation in living cells." Journal of the American Chemical Society 126.19 (2004): 5982-5983. PubMed PMID:...

...several ways. Below is a table listing some of critical differences (adapted from Zhou and Melton, 2008...this process. Moreover, it is much easier to genetically modify cells during the reprogramming process...This opens up a wide range of possibilities in clinical situations. In cases where the objective is to...Kushi) Mukherjee is the Editor of Trends in Pharmacological Sciences, a Cell Press reviews journal. She...from Massachusetts General Hospital and Harvard Medical School. Connect with her on LinkedIn @ https://...PMCID: PMC2854424. 23. Burridge, P.W., et al., Chemically defined generation of human cardiomyocytes. Nat...pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A, 2012....

...systems. FPs have been used to track protein localization, cell structure, intracellular trafficking, ...tagged proteins can lead to improper protein localization, protein aggregation, or disruption of normal...Also, it remains challenging to screen for the genetically modified strains that contain your newly tagged...recently developed a modular plasmid assembly toolkit called SapTrap to improve CRISPR genomic tagging in C....Protein Tagging. Beyond simply monitoring protein localization and activity, CRISPR tagging systems can be ...endogenous proteins for studying protein expression, localization, and protein-protein interactions. Additionally...

...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology...has facilitated the development of a simple and scalable means of making targeted genetic edits. Cas9 is... a gene of interest, they have been shown to dramatically alter levels of expression. Cas9’s ability to...Lance-Byrne is a graduate student at the University of California, Santa Cruz, where her research is focused on...Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease." Nature biotechnology ...Options for CRISPR Genotyping Cas9 Activators: A Practical Guide Additional Resources on Addgene.org Check...

...domain fragment derived from a camelid antibody called a heavy chain antibody (HCab). Because of their...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...enables rapid selection of synthetic nanobodies, called sybodies, against membrane protein targets in as...with a large and diverse pool of sybodies that hypothetically contain ones that bind strongly to your target... identifying sybodies that bind strongly and specifically to the protein of interest. The Seeger lab used...van Schie L, Hoffmann M, Pöhlmann S, Graham BS, Callewaert N, Schepens B, Saelens X, McLellan JS (2020) ...

...protein must be expressed (inducibly, ectopically, stably… it’s your call). The TIR1 F-box protein will associate...their fused protein under certain conditions (typically the addition of an exogenous small molecule). ... size from several kDas up to ~35 kDa and are typically knocked-in via gene targeting or cloned in-frame...proteasomal degradation. Adapted from Nabet et al., Chemical Biology, 2018.    HaloTag The HaloTag was ...protein structures, interacting partners, and localization. Thankfully, this is a hot topic with many resources...

...antibodies for every potential new target and basically impossible if you wanted to distinguish between...C-terminus post-translationally, which turned out to be critical for antibody recognition. As such, the peptide...interest and proteins tagged with it needed to be chemically treated before the antibody could detect them...investigating mechanisms of endoplasmic reticulum localization, Munro and Pelham casually introduced the Myc-tag... recognized by its antibody without additional chemical modification steps and when the epitope was within... of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable regions ...

... editing. Specifically, these types of CRISPR systems work using a multi-Cas complex called Cascade (CRISPR-associated... Cas3. Cas3 can induce genomic alterations on a scale that other Cas effectors can’t, while maintaining...recruit Cas3, which carries out the brunt of the mechanical work for editing. Class 1 CRISPR systems make...unraveling the double-stranded DNA (dsDNA) in what is called a reeling mechanism (see next section for details...debilitating disease! This is, of course, only hypothetical at this point. More time and energy must be ...

... the pharmaceutical industry for years. They have proven very difficult to target therapeutically due ...however, have yet to slay the beast. On a global scale, tens of millions of new cancer cases arise and ...fit. Kevan Shokat’s group at the University of California, San Francisco, is approaching the Ras problem... cancers, and synthesizing compounds to bind specifically to their mutated residues. For Ras researchers...represented, making the results of any such study as clinically relevant as possible. Connecting the research...

...delivery methods below: Chemical transfection   Lipid mediated Cationic polymers Calcium phosphate Cells... some CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator... be constitutive or inducible. U6 promoter is typically used for gRNA. May contain reporter gene (e.g....Medium Medium Medium Difficulty Easy Easy Easy Physical transfection   Electroporation / Nucleofection...

...is certainly not the first means to carry out so-called “forward genetic screening experiments”, it is ...in a particular phenotype. CRISPR can be readily scaled up for genome-wide screening due to the broad range...experiments you use them for! A well-developed biological question and experimental system are absolutely... to skewed results. Selecting a cell type - Theoretically, any cell type can be used in a CRISPR screen...screens check out Addgene’s 2016 publication “Practical Considerations for Using Pooled Lentiviral CRISPR...McDade JR, Waxmonsky NC, Swanson LE, Fan M (2016) Practical Considerations for Using Pooled Lentiviral CRISPR...