...gain-of-function screening when combined with a compact pooled sgRNA library. Our SAM-mediated screens exhibited...gene activation in mind. “We had not managed to create a system to allow us to reliably activate essentially...the next step in high-throughput, genome-wide screening,” Konermann said. “If you want to activate all...and make sure it’s working. You can only do that screen with a system you are really confident about working...her colleagues demonstrated the new system by screening for genes that, when turned on, make melanoma ...CRISPR library enabling genome-scale, knockout screening). As the Zhang lab's Nature paper reports, “Here...validation of the top 10 hits.” Two-way CRISPR screening In October, Jonathan Weissman’s lab introduced...

...DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre (FLP-dependent CRE) 55634  AAV1  pAAV-EF1a-mCherry-IRES-... constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-...membrane-embedding voltage detector. Find Archon1 and its cre-dependent version at Addgene! Fluorescent protein...

...these issues by creating immortal antibody-producing cell lines (hybridomas) that secrete antibodies in ...Neurotechnologies® (BRAIN) Initiative, will allow Addgene to create a much needed open-access recombinant antibody/...School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...reagent batch. To supplement this, Addgene will be creating a crowdsourced Data Hub (built off of the concept...resource to life,” says Dr. Trimmer. But in order to create such a resource, we need your help. You can participate... Director. “NABOR provides a new model for the creation and acquisition of affinity reagents and will ...funding laboratories, such as Dr. Trimmer’s lab, to create recombinant antibodies for use in neuroscience ...

...tested iTOP in a series of primary cell types using Cre and various loxP reporters. In both mouse and human...potential application of iTOP Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to...protein expression has a long-lasting effect, such as Cre- or Cas9-mediated recombination. Both primary hESCs... cre-lox...

...researchers injected Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent IL-1...injected AAV2 particles into a Drd1-Cre transgenic mouse to deliver Cre-dependent expression of  Channelrhodopsin...-1 beta overexpression cassette. Expression of Cre in these neurons turns on overexpression of IL-1 beta...2014). Scientists from the University of Geneva (Creed et al., 2015) used optogenetics to support these...transmission from the mPFC projections to the NAc and decreased the locomotion of the animal. This confirms the...behavior in rats in the forced swim test as seen by increasing the ratio of time spent actively swimming versus...described here is only a small selection of the increasing number of studies using viral vectors in behavioral...

...Expression of hM4D(Gi)-mCherry from this virus is cre-dependent, meaning that once injected, the DREADD...DREADD will only be expressed in cells that express cre recombinase. Typically, this feature is used to attain...cell-specific expression in transgenic animals, where cre expression is restricted to a particular cell type...the first time, the Roth Lab recommends verifying cre expression prior to use. The lab additionally recommends...expressing cell lines, which can then be used both for screens (using pooled CRISPR knockout libraries) and individual...Secondly, you can use a LentiCas9-Blast viral prep to create a single stable Cas9-expressing cell line and then...monoclonal Cas9-expression line prior to performing a screen or even a single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D...

... For example, a researcher can deliver Cre-dependent TVA to Cre-expressing neurons in a transgenic mouse...Addgene Blog AAVs CREATed for Gene Delivery to the CNS and PNS Advanced Uses of Cre-lox and Flp-FRT - ...viral protein EnvA on its surface will only infect Cre-positive, TVA-expressing cells (Osakada et al., 2013...

... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from...were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell...) alone at 1.1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct...

...interneurons can be labelled with an AAV1 Cre virus combined with a Cre-dependent AAV9 tdTomato virus in the...learn more about the AAV Data Hub, and hear the incredible AAV talks given by Connie Cepko and Ian Wickersham...

...Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering in species like... Plasmids 101: The Promoter Region Plasmids 101: Cre-lox Light-Switchable Two-Component Systems for E....Tetracycline (Tet) On Expression Systems at Addgene Cre-lox Guide and Plasmids     ...cell growth media with glucose decreases cAMP and represses pBad, decreasing promoter leakiness.   Other ...

...control when you turn off expression for good, the Cre-Lox system offers inducible yet permanent gene excision...can be excised by intentional introduction of the Cre recombinase. This is a time-tested system but does...some permanent knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for ...!  Turn it on Turn it on forever Permanently increasing or turning on expression of a new protein is ... gRNA, to your gene of interest to temporarily increase expression. There are several flavors of activators...

...labeling Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9...-4.   Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) by: Brian...multiplexed version of the Cre-mediated AAV Targeted Evolution method (M-CREATE), in which one applies both...variants that display increased neuronal expression in the brain while having decreased expression in the ...zebrafish as a model organism, the Parton Lab recently created a flexible system that makes use of pre-existing...Image from Goertsen D, et al., 2022.  Find M-CREATE plasmids here!  Goertsen D, et al. Nature Neuroscience...

...doxycycline, but it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular...Averof lab. It’s an alternative to the conventional Cre or Flp recombinases used to excise markers. It has...Cas12a inhibitors Using a bioinformatics and a screening approach, the Jennifer Doudna lab identified three...

...open-reading frame (DIO). Upon coexpression with cre recombinase (either from a mouse line or viral co-injection...type specificity dependent on the promoter of the cre recombinase while still taking advantage of the robust...an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...tracing with fluorescent proteins in nontransgenic and cre driver mice. Current protocols in neuroscience / ...: PMC4242517.  7. Madisen L, et al. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced... us to show causality in neural circuits with incredible temporal and spatial precision (I’m not going...optogenetics has already been proven. Therefore, a concrete, well defined, and robust behavioral output is...

...often made Cre-dependent. If Cre-dependent vectors are used in transgenic animals that express Cre in only...labelling the subset of infected cells that also express Cre. Another advantage of AAV vectors is that there are... traced neurons with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection into a conditional...conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the hippocampal...PMID: 4107258. Lo, L., and Anderson, D.J. (2011). A Cre-dependent, anterograde transsynaptic viral tracer...

... short hairpin) and CRUSH (Conditional RUSH) use Cre-mediated recombination to turn on or off the expression...a dual-color RNAi “sensor” mouse strain in which Cre expression causes the induction of both DsRed fluorescence...Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown in eyes and... recombination events at the genetic level. Upon Cre/loxP recombination, each transgene expresses one ...Flp recombinase/FRT system in place of Brainbow’s Cre/loxP, allowing for simultaneous use of the two systems...SUMO tags in the FP sequence for separation from Cre-based Brainbow-expressing cells. Autobow plasmids... all-in-one versions which express self-excising Cre recombinase from the same transgene as the Brainbow...

...sensors include syn-jYCaMP1s (slow variant), the Cre-dependent syn-FLEX-jYCaMP1s, and the axon-targeted...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...Find this AAV at Addgene New serotypes for the Cre-dependent Flpo recombinase pEF1a-DIO-FLPo-WPRE-hGHpA...biotechnology tool to optimize another. The authors have created a yeast-based directed evolution approach with ...your studies.  Find the plasmids to set up your screening systems at Addgene! Schematic of the evolution...following a DNA miniprep protocol!  First, the authors created the pMAK backbone by modifying pMAL-T-Avi-His/BirA...exported to the periplasm of bacteria where it can be secreted into the supernatant. Four plasmids containing...

...CI.mCerulean.WPRE.RBG James M. Wilson Cre Recombinase AV-1-PV1090 105537-AAV1 pENN.AAV.CMVs.Pl.Cre.rBG James M. Wilson AV....HI.eGFP-Cre.WPRE.SV40 James M. Wilson AV-1-PV2004 105545-AAV1 pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 James M....pENN.AAV.CamKII 0.4.Cre.SV40 James M. Wilson AV-1-PV2676 105553-AAV1 pENN.AAV.hSyn.Cre.WPRE.hGH James M. Wilson...-AAV1 pENN.AAV.CMVs.Pl.Cre.rBG James M. Wilson AV-2-PV1090 105537-AAV2 pENN.AAV.CMVs.Pl.Cre.rBG James ....HI.eGFP-Cre.WPRE.SV40 James M. Wilson AV-5-PV2004 105545-AAV5 pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 James M....pENN.AAV.CamKII 0.4.Cre.SV40 James M. Wilson AV-5-PV2408 105550-AAV5 pAAV.GFAP.Cre.WPRE.hGH James M. Wilson...-AAV6 pENN.AAV.CMVs.Pl.Cre.rBG James M. Wilson AV-8-PV1090 105537-AAV8 pENN.AAV.CMVs.Pl.Cre.rBG James ...

...in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions...community members. In addition we’ve experienced incredible growth with our readership expanding from ~10,000...next revolution in biological research. We’re incredibly happy to see that one of our career posts has...protein of interest. 4. Plasmids 101: Blue-White Screening Getting a lot of empty vector contaminants in ...Addgenie Jessica Welch explains how Blue-White screening could be a potential solution. 5. Cpf1: A New ...

... the expression of a Cre recombinase contained within the knock-in. Including Cre-mediated marker excision...kilobases of additional sequence being introduced. Cre-mediated excision of the selective cassette minimizes...experimental manipulation. In some cases, injection of Cre is required, however the development of the self-...variable amounts of experimental manipulation and screening.  A co-selection strategy for CRISPR editing in... with inefficient sgRNAs in my initial direct screening efforts, which were similar to the approach detailed...low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering rare editing events...conditional-lethal mutation to identify edits with minimal screening effort. Selecting for repair of a lethal mutation...