...activity to a transcriptional activation domain (Fig 1). Two ways to use CRISPRa for your research CRISPRa... Stucka R, Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional... (2014) A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging.....org Find Cas9 Activators Find Cas9 Repressors Catch Up on Your CRISPR Background with Our CRISPR Guide...

...breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime editing. Created with BioRender.com...demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations have continued...Engineering. https://doi.org/10.1038/s41551-024-01227-1 Yan, J., Oyler-Castrillo, P., Ravisankar, P., Ward... the indicated tool. ~ reflects an innovation that may optionally be combined with the indicated tool....genetic variants have been identified in humans and cataloged in the ClinVar database. Previously developed ...prime editing Less constrained by PAM sequence location The prime editor extends the reach of CRISPR genome...editing typically needs to be optimized for each application, and the efficacy varies widely — it still almost...

...is here to help. Contact us at [email protected] or +1 617-225-9000, or check out our MTA reference pages...results, modifications, and inventions created by the Recipient Right to file patent applications on modifications... about intellectual property rights, pending publications, and commercial use often come up. An MTA can...trials Acknowledgement in recipient’s future publications relating to the material Limited liability...

...Cloning grade DNA Transfection grade DNA Figure 1: Transformation introduces transformation grade plasmids...that have been used during the purification procedure. Good indicators of DNA purity are the absorbance...nm should fall ~1.8 - 2 and give scientists an indication of purity versus protein contaminants. The 260...nm ratios should fall ~2 - 2.2, and provide an indication of purity versus chaotropic agents as guanidine...

... successful after their training.   Figure 1: Professional Development Hub Workshop Attendees and... It's been an honor to work with such a group dedicated to providing the structure and know-how to integrate...example, pd|hub assembled a coalition of higher education organizations committed to advancing career and... inclusive career readiness training in this educational path before we lose more scientists who might...

...’ve come to the right place. Get started with PGC-1 plasmids from Addgene. References: Skeletal muscle...window onto communication between the muscle and brain, which may have therapeutic implications, say Hyo ...

... with DNA sequencing troubleshooting and analysis 1. You can use any of the following programs to view...of interest. 5. Use a silica spin column for purification of the samples you send for DNA sequencing If...requires you to perform your own Big Dye PCR amplification reaction (as opposed to using the all inclusive...

...Histochemistry & Cytochemistry, vol. 59, no. 1, SAGE Publications, Jan. 2011, pp. 6–12. Crossref, doi:10.1369...compare the experimental group.     Figure 1: A, Most biological experiments will benefit from ...results.  Replicates Replicates are another critical control in biological experiments. Replicates can be...of biological replicates such as multiple mice in each group and C, technical replicates or repeated measurements...secondary antibodies.   Application-specific controls Some antibody-based applications have very specific ...then moving on to controls for antibody-related applications. After reading, you will have a better understanding...blot and look for the molecular weight shifts indicative of tau phosphorylation. By including the positive...

...science’s place in education, government, and civilization in general (1). Science and politics? Science...consequences of a march. Three of these risks include: 1) Lost trust Perhaps the greatest risk is that the ...sister locations on April 22, 2017, scientists and non-scientists alike will march to advocate for science...already come to pass. It may only be by advocating for science education and literacy that this changes.  2...marching for. I will be marching to advocate for science education and literacy as I believe people that...communities, microbial interactions, and science education.  Disclaimer: The views presented in this article...process begins with a testable hypothesis - an educated guess about how some part of the world functions...

...improved sensors for these and other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted...fluorescent biosensors. Nat Commun. 2022 May 25;13(1):2919. doi: 10.1038/s41467-022-30685-x. Feel the ...sensors One of the latest additions to Addgene’s catalog is a new class of sensors for neuropeptides and...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...They generated variants optimized for different applications (v82 has higher affinity for glutamate but slower...PDGFR.codonopt    Aggarwal A, et al. Glutamate indicators with improved activation kinetics and localization...have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized...

...detects the light scattered by that cell at a low angle (FSC) or large angle (SSC) (Figure 1). FSC values ...colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered forward at a low angle... you with two measurements: forward scatter (FSC) and side scatter (SSC). To acquire these measurements...exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably...types of scattering that occur when a cell is pulsed by the laser. Within each type of scattering, you also...to keep it short though, promise! FSC and SSC: scattering To digitally extract your cells based on their...passes through the laser beam and causes it to scatter (FSC/SSC), the detected photons can be represented...

...together along with the Gibson assembly master mix for 1 hour, and then transform into competent cells. The...you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison...neighboring fragments can be created via PCR amplification with primers that contain the appropriate homologous... tools that help you design primers for PCR amplification of fragments to incorporate such regions of ...assembly meets CRISPR Gibson can be adapted to more complicated cloning schemes, such as those where the vector...CRISPR technology and is described in a recent publication by the Lockey lab (Wang, et al. 2015). In this...aspects of the SLIC methods. First, it uses a dedicated 5’ exonuclease instead of using the exonuclease...

...example, if there are 10 putative off-target locations you can 1) determine if editing at those sites would...downstream experiments. You only successfully generate 1 null clone and proceed with it. In this case, the ...“Off-target effects” a vague catch-all term for unintended consequences in an experiment. For CRISPR ...sequencing. While this may be unnecessary for many applications, it is the only comprehensive method available...binding by Cas does not always correlate with catalytic activity at those sites, especially when DNA mismatches...For a candidate site-specific approach, mRNA quantification or western blot of putative off-targets would...you’ve tried everything listed here. For some applications, this is ok, but it still must be controlled...

...out these SciComm blog posts. References 1. Bourdeau, Raymond W., et al. "Acoustic reporter genes for... science communication and outreach - if you'd like to learn more about Science communication check out...microscopes and give a similar optical quality (magnification of 140X and 2 micron resolution). I have worked...creative approaches to visualize bacteria in an educational setting. The Symposium on ‘Creative Approaches...from hand-drawn comics (Dr. Sanja Saftic) to educational gifs created by the Amoeba Sisters. Here’s one...Bacterial Genome Engineering Learn about Science Communication using Twitter  Resources on Addgene.org Check...

... remains fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained in...follow him on twitter: @joachimgoedhart.   References 1. Zacharias, David A., et al. "Partitioning of lipid-modified... proteins.Therefore, GFP is not suitable for applications that require anaerobic conditions. In experiments...an additional protein module that can report on location. This approach enables the study of proteins in...fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps...case when proteins undergo post-translational modification with fatty acyl chains. For instance, myristoylation...C-terminal consensus sequence (-CaaX). Both modifications are disrupted when GFP is fused to these sequences...

...five methods for extracting DNA without a kit too. 1. Early Career Researcher Toolbox: Free Online Molecular...virtually. 6. Quantifying DNA? Here are Five DNA Quantification Methods to Consider by Jennifer Tsang The...The nanodrop vs. spec debate dominates DNA quantification discussions, but what about the other methods... with plasmids deposited at Addgene! 2. DNA Purification Without a Kit by Beth Kenkel To complement ...

... two years. References 1. Martz, Colin A., et al. "Systematic identification of signaling pathways with... of 17 signaling pathways that are frequently implicated in cancer cell proliferation, survival, differentiation... make the library usable for pooled screens. Applications of the cancer pathways ORFs kit Wood and his...

...infections, cancer, and other diseases. References 1. Harrington, Lucas B., et al. "Programmed DNA destruction...of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this viral sequence in the assay...

...regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed.  Related Resources...says the new tool might even find therapeutic application in addition to their use in basic biology, for...

...to the interview with Science in the News Chapter 1: Introduction - Vini and Amy tell us a little about... to be crazy complicated! Listen to the full podcast for more great science communication tips or listen...tons of ways you can get involved in science communication. In this second half of our conversation with...many things you can do start your own science communication student group and get more involved with your...is a graduate student group with the goal of communicating science to the general public and training the... the next generation of science communicators. In this chapter we learn about some of their events - science... peer-to-peer feedback in effective science communication. Chapter 7: The Goal of the SITN Podcast - Vini...