...? Depositing lab: The name of the Principal Investigator (PI) in whose lab the construct was made. The...

...Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been ...developing and depositing two new norepinephrine indicators: nLightG and nLightR (Kagiampaki et al., 2023...White insets indicate surface expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T...trialed in two-photon in vivo experiments. nLight indicators are more sensitive to and selective for norepinephrine...They can, however, be studied with the nLight indicators. Speed One of the major advantages of this next...… Patriarchi, T. (2023). Sensitive multicolor indicators for monitoring norepinephrine in vivo. Nature...

...Voltage indicators are voltmeters for cells! In this blog we will review how these indicators work, their...fluorescence or voltage indicators.   Viral Vectors and Voltage Indicators As previously mentioned...their use as biological tools. Why Use Voltage Indicators? Electrical signals are integral to many facets...within a cell are genetically encoded voltage indicators (GEVIs) and chemigenetic sensors. These tools...describe in more detail below. Types of Voltage Indicators GEVIs can be divided up structurally into 2 main...structurally separates the two components of the indicator, which can make the kinetics of voltage sensing...Factors like photobleaching, lack of red-shifted indicators, and brightness of fluorescence all have room...

...plasmids use Rho-independent terminators, which include naturally occurring terminators, such as T7 and rrnB,...engineered terminators come close (>95%). For most purposes, however, any common terminator will suffice...double terminators to reduce unwanted translation of downstream elements. A high affinity terminator may ...of yeast terminators of varying strength. A discussion of the effects of various terminators in the context...termination and polyadenylation? The role of the terminator, a sequence-based element, is to define the end...synthesized RNA from the transcription machinery. Terminators are found downstream of the gene to be transcribed...transcribed, and typically occur directly after any 3’ regulatory elements, such as the polyadenylation or poly...

...California Institute of Technology Cold Spring Harbor Laboratory  Dana-Farber Cancer Institute  Fred Hutchinson...

... download the PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging ...corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier to plan...for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting...right, you need to do a few calculations. This calculator can do the math for you and it’s simple to make...whole experiment And here’s the equation the calculator uses to give you the μl needed of each plasmid...experiments. Let us know what you think about the calculator in the comments section below. References 1.... transfection calculator...

...with us. Tom McFadden is a YouTuber and science educator looking for creative ways to implement the next...

...isoforms, to help researchers understand this regulatory network. The transcription atlas and library...deliver multiple TFs to each cell, allowing for combinatorial overexpression programs on cellular processes...

... an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your favorite biological...screening are vastly useful for mapping the combinatorial genetics that underlie complex regulation in...barcoded gRNA libraries that can be used to combinatorially modify the genome, screen for a particular ...successfully applied to functionally characterize combinatorial gene knockouts generated via multiplexed gRNA...expression (Wong et al., 2016), in addition to the combinatorial expression of other genetic elements including... Figure 2: Strategy for assembling barcoded combinatorial gRNA libraries. (Adapted from Wong et al., 2016...antibiotic-resistant bacteria enabled by massively parallel combinatorial genetics." Proceedings of the National Academy...

...we introduce you to the Journal of Emerging Investigators, an open-access journal that enables high school... the interview with the Journal of Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction...Olivia Ho-Shing, and the Journal of Emerging Investigators. Chapter 2 - Getting high school students and...

... plasmid toolkit, the Broad Institute Target Accelerator Plasmid Collection, that we hope researchers ... For many years, we’ve hoped that large scale laboratory-based functional experiments that classify which...and cancer phenotypes. Broad Institute Target Accelerator Plasmid Collections To overcome this first challenge..., we’ve developed the Broad Institute Target Accelerator Plasmid Collections, now widely available to ...Collection). In addition, we worked with many collaborators to aggregate a sequence-confirmed series of ...

...School and Paratore is the General Auditor at Tufts Health Plan. The addition of Katz and Paratore onto the...Interactive. Learn more about Carl Paratore, MBA, CPA Mr. Paratore serves as the General Auditor at Tufts...re excited to welcome Karen Katz, Esq and Carl Paratore, MBA, CPA to the Addgene Board of Directors. Katz...that Addgene was looking for new board members. Paratore: Kristen Darby, who’s also a board member, reached...necessary information that’s required to do research. Paratore: What I’ve been most impressed with is the level...potentially also be very helpful to the scientific community.Paratore: I’ve certainly had a front row seat watching...be, an interesting legal and professional challenge.Paratore: The promise there is the ability share the...

...potent transcriptional activators (p65 and Rta). These transcriptional activators work in tandem to recruit...CRISPRa methods vary in their transcriptional activators: some methods rely on fusion proteins while others...generally thought of as the “first generation” CRISPR activator. While it requires a relatively simple construct...modest gene activation. Synergistic Activation Mediator (SAM) Description SAM uses specially engineered...levels of gene activation compared to other CRISPR activators, making it a popular method for gene activation... SunTag performs better than first generation activators while showing lower activation levels than SAM...transcription. This complex consists of the VP64 activator used in other CRISPR activation methods, as well...

...voltage indicator Archon Article contributed by Shreya Vedantam  Listen to the voltage indicator podcast... genetically encoded voltage indicators. To expand the voltage indicator toolbox, Ed Boyden’s lab developed...Archon1, a novel opsin-based fluorescent voltage indicator. Archon1 can embed itself into the cell membrane...

... for a curated list of coregulators). Coregulators can function as coactivators and corepressors depending...complexes with secondary coregulator (or co-coregulator) partners. The coregulator ‘core’ complex is composed...feature afforded by coactivator complexes suggests that coactivators may be integrators of multiple cell ...illnesses, and the metabolic syndrome. Coregulators Coregulators constitute a group of some 400 molecules...that coregulators belong to distinct complexes may explain how more than 300 different coactivator proteins...manner. Because coregulators exist as multiprotein complexes, a member of a single coactivator complex can... W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators. Cell 108, ...

...Choose a moderator with a backbone. It is nice to have a subject area expert, but the moderator is not ...LinkedIn profiles of all panelists to the moderator. The moderator and organizers should develop a list of...(lapel mics are best). The moderator takes control It is the moderator's job to prevent anyone from ...effortless to the audience, but the organizers and moderator should have a well-planned approach to making ... of 4-6 pre-event interview questions. The moderator (possibly with an organizer) should set up a 30 minute...to schedule. In the pre-event interviews, the moderator should be taking good notes and discovering interesting...opinions that each panelist can contribute. The moderator should identify the funny, provocative or interesting...

... the perfect voltage indicator and optogenetic actuator pair. Ideally, indicators should possess fast ...lab wished to improve on the Arch indicator and their collaborators in Robert Campbell’s lab carried out...kinetics. By combining both the voltage indicator, QuasAr, and actuator, CheRiff, in the bicistronic Optopatch...activation of specific genes using dCas9-VP64 activator fragments (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(...62885 & 62886) Rapamycin-inducible dCas9-VP64 activator set (Addgene plasmids 62887 & 62888) Zetsche... which will ultimately co-localize the VP64 transactivator with a dCas9 that has been targeted to a very...tool for activating endogenous genes Moritoshi Sato's Lab has engineered CRISPR-Cas9 to create a photoactivatable...

...Roth New serotype pAAV-EF1α-F-FLEX-mNaChBac-T2A-tdTomato AAV8 Molecular tool Massimo Scanziani New viral...Optogenetics Jonathan Ting New viral prep pAAV_BiSSTe10_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New...Optogenetics Gordon Fishell New viral prep pAAV_BiCHATe27_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New...Optogenetics Gordon Fishell New viral prep pAAV_BiVIPe4_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New...Gordon Fishell New viral prep pAAV_BiLAMP5e3_dTomato_nlsdTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell...Optogenetics Gordon Fishell New viral prep pAAV_BiPVe4_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New...Optogenetics Gordon Fishell New viral prep pAAV_BiPVe3_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New...

...cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based research as...of the gene, Cas9 activators efficiently solve these problems.   Which Cas9 activator should I use? There...significant work on constructing Cas9 transcriptional activators. These constructs allow for the upregulation ...genome-wide studies One of the best uses for Cas9 activators is in genetic screening. gRNAs targeting every... using oligo library synthesis. Prior to Cas9 activators, similar tools were made using other DNA binding...to easily change the sequence targeted by the activator by simply providing a new gRNA rather than engineering...organism, have been used in a similar manner to Cas9 activators. However, these can be difficult to construct...

...Repressible: An activator protein is bound, turning transcription ON. When a repressor binds the activator protein...introduced the second generation QF2 activators. These activators solve the problem of broad QF expression-induced...protein, the activator protein cannot bind the promoter sequence anymore and transcription is turned OFF....protein, the repressor protein can bind to the operator. The bound repressor then prevents transcription... repressor (TetR) can bind to the tetracycline operator sequences (TetO), preventing transcription. In... Bujard created the tetracycline-controlled transactivator (tTA). They fused TetR with the transcriptional...by Gossen and Bujard, both the promoter and transactivator components of the system have been engineered...