...technology, the Vale and Weissman labs took this biological lesson and created a system to amplify fluorescent...modified the GCN4 sequence to increase its alpha-helical structure and stability, creating v4 SunTag. Since...aggregation, it’s recommended for most imaging applications. What can you do with SunTag? Tanenbaum et al...flames,” it’s clear that the system is broadly applicable across many areas of research. Beyond fluorescence...activation of target genes, but we’ll save that application for another day! References 1. Tanenbaum, Marvin...et al. “A protein-tagging system for signal amplification in gene expression and fluorescence imaging....

...generally linked to dopamine (DA), a well-known reward chemical and neurotransmitter. But aside from DA’s famous...at a certain frequency, and this frequency can indicate behavioral state, for example, if someone is awake... it can be performed in a live awake animal by surgical implantation of the probe into a specific brain...in freely moving mice Record DA dynamics of optogenetically activated and inhibited neurons in live mice...neuronal imaging of dopamine dynamics with designed genetically encoded sensors." Science 360.6396 (2018): eaat4422... PMCID: PMC6287765. Sun, Fangmiao, et al. "A genetically encoded fluorescent sensor enables rapid and ...

...swimming where they periodically beat their tails left and right in a symmetrical motion to move forward...behavioral studies. In neuroscience, opsins are typically ion channels that allow charged particles to cross...Kolmer-Agduhr cells was sufficient to trigger the symmetrical tail movement unique to spontaneous slow forward... photostimulation in order to drive the full symmetrical response.  The function of Kolmer-Agduhr cells...Aristides B., Filippo Del Bene, and Herwig Baier. "Optical control of zebrafish behavior with halorhodopsin...

... (e.g. 6xHis containing pDEST-HisMBP). After purification the protein can be released from the resin and...types of yeast expression vectors. Consider your applications and experimental design. You can create a new...more traditional approach of an autonomously replicating vector that carries either a wild-type version...expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern that allows the ...a variety of other types of vectors for most applications used to study gene function. From making transgenic...transgenic animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene ...

... Foundation Round Table. In any field, good communication skills help when it comes to finding a job. ...an argument for taking the time to hone your communication skills. So, where can a graduate student or ...opportunities than ever before for scientists to communicate about their work and their interest in science...your area or at your institution; the news or communications office is a good place to start. It’s not unusual...about science writing as a career, the Science Communication Program at the University of California, Santa... your research stories. Consider practicing communicating your research by guest blogging here, just leave...

...problems, develop general technical proficiency, communicate your work, maintain ethical standards and interact...? Listen to our podcast interview with Harvard Medical School researcher Connie Cepko to learn about her... you admire her management style? Does he have ethical behavior? Does the lab encourage diversity? Knowing...websites listing alumi and it is easy to use publication records, university websites, ResearchGate and...

...appreciable levels of modification. Whole-genome sequencing of cells modified by CRISPR indicates that the consequences...20nt-NGG] and [5’-CCN-20nt], as it is equally efficacious to target the coding or non-coding strand of ...desired location of edit; efficiency decreases dramatically when the cut site is >30nt from the proximal... via CRISPR cutting. For CRISPRa, it is most-efficacious to target a ~100nt window upstream of the transcription...sequence may not be available. Predicting gRNA efficacy We and others have examined the ability to use...needs to test in the lab. Importantly, for any modification of interest, it would be unwise to make conclusions...

...natural and engineered repressible promoters. Chemically repressible promoters Tet-Off In its native context... GAL80 under different promoters and create sophisticated patterns of UAS-driven gene expression. To further... PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic... PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, Christopher J. Potter... PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, et al. “Improved and ... PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O, Potter CJ. The Q-System...expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res...

...the gRNA design process and rules for various applications is necessary going forward. The standardization...benefits to researchers conducting basic or early pre-clinical research. However, standard terms and techniques... I think I may have finally grasped just how complicated DNA repair pathways are. I fully admit to being...to me. But coming to #KSgenome and seeing the intricacies of DNA repair has emphasized how essential these...Alt-NHEJ), has been harnessed for CRISPR knock-in applications. CRISPR 101: Targeting RNA with Cas13a (C2c2...for CRISPR Genome Editing Other resources and publications CRISPOR gRNA design tool Researchers at the ...

...supercoiled DNA ends to facilitate replication. The TOPO technique specifically uses Vaccinia virus-isolated...insert comes from using Taq polymerase for the amplification step since Taq polymerase leaves a single deoxyadenosine...minutes (incubating the full 30 minutes will not significantly improve the transformation efficiency).  5. ... has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading functionality are...are planning on transforming your reaction into chemically or electro-competent E. coli - excess salt causes...

...University and Medical Center  Weizmann Institute of Science Whitehead Institute for Biomedical Research ...time of request MTAs and deposit agreements to basically instantaneous in many cases.  Suggest that they...your organization below? Don’t worry--we check periodically for possible new awardees, and we’re happy to...University Rockefeller University  Salk Institute for Biological Studies The Scripps Research Institute St. Jude...Massachusetts, Worcester  University of Nebraska Medical Center University of Pennsylvania  University of...

...Addgene! Archon1, a voltage sensor AAV Historically, electrical activity in the brain was measured with...kinetics) that makes them suitable for different applications. We recently released a new variant of jGCaMP7... been developed and are suited for different applications. The iGABASnFR GABA sensor (pAAV.hSyn-FLEX.iGABASnFR...voltage reporter allows a way to monitor neuronal electrical activity through imaging. Since it’s illuminated...Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...

...infection, because this setup allows for experimental replicates for each of the gRNAs. The idea was to assign...genetic tool for labeling just a fraction of a genetically-defined subset of cells. By labeling fewer cells...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ...measurements, or monitoring in vivo. To overcome these technical issues, Sabrina Büttner’s lab designed a calcineurin... reporter (blue) and stained with PI (pink) to indicate cell death. Image from Diessl et al., 2020. ...protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison...

... (AmpR) and origins of replication (ori and f1 ori) which allow it to replicate.   How can I prevent...the bacterial selection marker and origin of replication. During bacterial culture, only the plasmid with...the antibiotic resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria...the recombined plasmid is still visible. This indicates that the DNA prep contains a mixture of the full...MK, Rudolph C (2019) Segmented poly(A) tails significantly reduce recombination of plasmid DNA without ...

... “species on species”) immunochemistry (IHC) applications an issue? The culprits are endogenous IgGs in...IgGs and your primary antibody, resulting in significant background staining (Figure 1A). Not all tissues...Figure 2B) Some labs have found that you can significantly decrease background staining by mixing your ...Randolph-Habecker J (2013). A Flexible Mouse-On-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free...Partridge TA (1998). A New Blocking Method for Application of Murine Monoclonal Antibody to Mouse Tissue...Tuson JR, Pascoe EW, Jacob DA (1990) A novel immunohistochemical technique for demonstration of specific binding...

...lab used an alternative strategy to develop a genetically encoded antibody based probe that detects the...antibodies for extracellular and intracellular applications: CDR grafting to stable frameworks and structure-based...National Academy of Sciences of the United States of America, 69(9), 2659–2662. PubMed PMID:4560694. PubMed ... and antibodies with crystalline papain. The Biochemical Journal, 73(1948), 119–126. PubMed PMID: 14434282..., T., Sato, Y., … Stasevich, T. J. (2019). A genetically encoded probe for imaging nascent and mature ...mature HA-tagged proteins in vivo. Nature Communications, 10(1). PubMed PMID: 31270320. PubMed Central PMCID...

...Harvard Medical School and runs everything science (CSO). Clinton is a cost analyst at Dow Chemical (a Fortune...a company in the health sciences without those critical three letters after your name (P-H-D).  Finally...two projects, including one led by a talented biomedical engineering team from MIT.   I would like to ...launching a company without partners.  You need a dedicated team that you know well and trust.  This will ...majority of these meetings acted as a platform for significant company advancement. Be relentless The world ...

...databases available, Web of Science. For each publication, Web of Science provides basic bibliographic ...Microbiology, Biochemistry & Molecular Biology, and Biochemical Research Methods. We also pulled the big interdisciplinary...Science, PLOS One, and PNAS. We then broke the publications from these journals into two groups: those containing...the papers with plasmids deposited at Addgene – typically about four times as many as papers without plasmids...differences in citations due to the journal of publication. Table 1 provides an example calculation of the...Addgene in them. The adjustment is done within publication years. What do we find once we do this adjustment...

...field has exploded so quickly, citing the first clinical trial with iPSCs with his collaborator Takahashi... have profound impacts on the development of applications with the potential to save lives. While most...picture perspective: Dr. Yamanaka talked about his clinical trial patients with advanced macular degeneration... ground-breaking work on spinal cord injuries; Jessica Young shared research on stem cells and Alzheimer's...Eggan presented work that is leading to promising clinical trials for ALS.   I have an uncle with Parkinson's...the first stem cell line that could be used in clinical trials. The quality control process alone takes...

...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...has been widely adapted for a wide variety of applications: base editing, CRISPR activation and inhibition...Pederson’s lab at the University of Massachusetts Medical School developed a dCas9-based multicolor labeling...RFP) to the genomic loci of interest. Pairs of identical hairpins within one gRNA will result in a blue...National Academy of Sciences of the United States of America 112 10 (2015): 3002-7. PubMed PMID: 25713381 PubMed...al. “CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging.” Nature Methods 15 (2018)...

... distancing? Many people are now calling this “physical distancing” since it’s important to stay virtually...message for some people. We should recommend #PhysicalDistancing. Social interactions and #social support ...for all ages. Organize your data and prepare publications or your dissertation Organize data that you...these drafts and for a few model organisms, Micropublication will accept negative data or publish brief...brief, novel findings. Most Micropublications contain one figure. If you aren’t there yet, write introductions...introductions to potential publications or prepare a review article. These are both options that can be done...

...Glutamate plays a variety of roles in synaptic communication and can trigger other forms of neuronal signaling...SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate Indicators for Synaptic Imaging However, the first two generations...researchers did the following: Improve sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic...membrane trafficking and localization. Improved photophysical properties The improved variants of iGluSnFR3...Blog Learn about other Fluorescent Biosensors A Practical Approach to Choosing the B(right)est Fluorescent... 30377363 Aggarwal, Abhi, et al. . “Glutamate indicators with improved activation kinetics and localiation...

... outcomes. The quality of the work is often dramatically improved when tasks are distributed appropriately...team manager must then facilitate appropriate communication between members of the team to ensure that no...of the overall project gets left undone or is duplicated. With each component of the project completed.... Listen to our podcast interview with Harvard Medical School researcher Connie Cepko to learn about her...clear outcomes in mind, you won’t be able to communicate these well to the task assignee and this is a...with most managerial responsibilities, good communication is at the heart of delegating for success.  ...

...plasmids can be used to carry out any biochemical and biophysical study involving protein stability or ...degradation. Despite how easy these experiments are to physically set up in the lab, RNAi is often completely ineffective... most scientists use mutant animals that are genetically altered to overexpress endogenous RNAi pathways...created an improved RNAi vector that leads to significantly stronger phenotypes than standard RNAi protocols...facilitate protein-protein interactions and mediate biochemical processes. One of the largest and best characterized...

...Stupid, overwhelmed, discouraged, tired…words that indicated there were many barriers between our audience ...readers.    Figure 1: An image from AAV Purification by Iodixanol Gradient Ultracentrifugation ...Figure 1 appears on our protocol page for AAV Purification by Iodixanol Gradient Ultracentrifugation. The... reader accessing the information? This can be physical or digital. For instance, you would present information...most helpful to use a lot of jargon and have a technical, detailed approach. Other times, it is more helpful...thought it would be most helpful to highlight the technical details of collecting the purified AAV–that the...

... useful information on scientific literature, technical skills, and experiences from thousands of laboratories...antibody and content team, we identified what applications should be included and what basic information... The guide was then assigned to our science communication specialist, Rachel, to write. Identify expertise...through an editing process and final check before publication.   Do research! Nearly everything we write entails...Guide figures. This requires a fair amount of communication between our content team and our graphic designer...graphic designer upon request that the science communicator "just sketch a western blot out quickly." Final...

... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be...1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011 - 1x1012 vg/...Have you or others been able to successfully systemically transduce and then retransduce a model organism...as multiple sclerosis?  A: Basic science and preclinical researchers have used the flexibility of AAV ...CRISPR tools is an exciting area for future AAV application to epigenetic editing. This area is discussed...

...cells and antibody-producing B cells that cannot replicate, meaning that only hybridomas will survive this...antibody, meaning that the antibodies are largely identical between lots.  Disadvantages of monoclonal antibodies...there will be changes between lots over time. Specifically, the antigen-binding sites of the antibody -... Antibodies: Distinguishing Characteristics, Applications, and Information Resources. ILAR Journal 46:...2016) Ligand-induced Epitope Masking. Journal of Biological Chemistry 291:20993–21007. https://doi.org/10.1074... the Addgene blog Learn more about research applications for antibodies in Intro to Antibodies Read about...

...getting to talk to so many different people! I communicate with customers, software developers, as well ...from interacting with others, and have strong communication skills to tailor your message to any type of... to solve is clear, but the solution is more complicated. You’ll need to be creative to be able to work...m always catching up in the morning! 9:00 AM--Technical implementation review. The developers discuss ...automated protocols. 12:00 PM--Catch up. Finish any communications with my UK team members before they log off... at Oxford Nanopore Technologies for the North America region. She is particularly interested in DNA sequencing...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ... department at the University of Massachusetts Medical School. We are working with postdocs at other institutions...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... December 1st. Salary inconsistencies across geographical regions When we originally started gathering...changes in science. Follow him on Twitter @BiophysicalFrog and Future of Research @FORsymp.   Additional...