... directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene through...pSpCas9(BB)-2A-Puro (PX459) V2.0 from it's predecessor, which had a point mutation in the puromycin resistance...most request plasmid in 2016 was... pSpCas9(BB)-2A-Puro (PX459) V2.0       Similar to last year, the top...through a 2A peptide. The puromycin resistance gene enables selection of cells with the plasmid post transfection...causes the ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This...mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro (PX459) V2.0: What's not in the name? Not included...

...., et al. (2023) Neuron, 111(10), 1547–1563.e9. doi: https://doi.org/10.1016/j.neuron.2023.03.009. PMID...microglia The REVeRT Dual AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation... vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.   ...fidelity during action potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage...vectors and are suitable for extended imaging of neurons in vivo. Because they are negative-going sensors... the vector to silence transgene expression in neurons and other non-microglia cells.      Figure...42386-0. PMID: 37852949. New and notable Neurodegeneration Collection updates Last but not least, check...

...continues to supply neuroscience with ever more efficient tools for light mediated neuronal control and we ...pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter sensors: Dopamine and norepinephrine Neuroscience research relies on...photostimulation of axons and dendrites on neighboring neurons. We have also packaged hChR2(H134R) plasmids from...calcium dynamics that govern such activities as neurotransmitter release is vital in studying neural networks... on accurate monitoring of neurotransmitters and we are excited to offer AAV for the newest GRAB sensors...tool from Loren Looger’s lab at Janelia for neuroscientists to measure glutamate levels. iGluSnFR.A184S...

...Oliver Griesbeck of the Max Planck Institute for Neurobiology has been working on genetically encoded indicators...monitoring the activity and biochemistry of living neurons in an effort to understand the connection between...considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors Griesbeck’s...available beam splitters. In addition to imaging neurons, Griesbeck adds that this color change has a particular...refined by a secondary screen in rat hippocampal neuron cultures. The optimized Twitch sensors allowed ...researchers to see tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes...them a versatile tool for application in both neuroscience and immunology. Don’t be shy: Ratiometric imaging...

...be used for sparse labeling of neurons for tracing experiments.  Neuronal tracing experiments: While synthetic...broad number of cell types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than ...that produce sparse labeling would be useful for neuronal tracing experiments where lower levels of labeling...labeling makes it easier to trace a neuron’s projections. Generating animal models: Transgenic animals allow...for retinal gene therapy Find more about precise neuronal targeting using AAV technologies Browse all of... H.P., Krol, J., & Roska, B. (2019). Targeting neuronal and glial cell types with synthetic promoter AAVs... mice, non-human primates and humans. Nature Neuroscience, 22, 1345-1356. https://doi.org/10.1038/s41593...

...pushed the boundaries in neuroscience by granting targeted control over neuronal activity. While they serve...scale of milliseconds, and can precisely link a neuron's activation to behaviors. Scientists continue to... mutated to have faster off kinetics and allow neurons to repolarize faster.  Chemogenetics does not offer...Administering the drug once can result in several hours of neuronal excitation or inhibition. This sustained activation...tissues, cell types or even subcellular regions of a neuron. Optogenetic approaches excel at discrete cell ...precise spatiotemporal control of a small subset of neurons is necessary, use optogenetics. If broad control...

... reagent resource for the neuroscience community called NABOR (Neuroscience AntiBody Open Resource). We...Brain Research through Advancing Innovative Neurotechnologies® (BRAIN) Initiative, will allow Addgene to...thousands of high-quality antibodies/nanobodies for neuroscience research and is an expert at validating these...While the focus of the NABOR platform will be neuroscience reagents and data, the platform can be expanded...are among the most commonly used reagents in neuroscience research today, but there is a critical unmet... to create recombinant antibodies for use in neuroscience research. More broadly, advances in gene synthesis...

...are powerful tools to modulate the activity of neurons and other brain cells. Since the opsins or chemogenetic... we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or mice (Galvan et al., 2019)... electron microscope showed us that, in monkey neurons, the large majority of the gold particles were ...found a larger proportion on the membrane in mice neurons, most of them were still cytoplasmic. Use of ‘small...hM4Di‐mCherry at the plasma membrane in monkey neurons, we next hypothesized that a different tag fused...instance, when the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells...Ultrastructural localization of DREADD s in monkeys. Eur J Neurosci 50:2801–2813 . https://doi.org/10.1111/ejn.14429...

...presynaptic region of one neuron and postsynaptic region of a different neuron, come into proximity in ...Druckmann et al., Neuron. 2014. Feb 5;81(3):629-40.  Next-Gen Brainbow toolkit for neuronal imaging Joshua...perform independent two-color optical excitation of neurons. The paper describes five previously unknown ...allowing labs to precisely and non-invasively control neuronal signaling. For more information on DREADDs, ...Addgene's Genome Engineering Guide. Fire up those neurons: mGRASP A Nature 2012 article by Jinny Kim and... use of another set of mGRASP plasmids in their Neuron 2014 paper. Kim et al., Nat Methods. 2011. Dec...high-resolution fluorescent imaging of individual neurons. The technology generates a unique spectral identity...

...norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype. ...challenge to scientists. What works well in one cell or neuron type may not work well for a neighboring cell type...it to publish!  Working with AAV vectors in neuroscience Let’s consider my first challenge when I started...express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes containing an expression...84%) were both excellent at infecting our target neurons (Figure 1). However, this one set of experiments...in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https://doi.org/10.3389...

...particularly well-suited for neuroscience investigations. For example, neuronal activity can be tracked in...available from strain distribution centers in the US, Europe, and China. In addition, the Zebrafish Model Organism...example, specific tissues such as blood cells or neurons can be fluorescently labeled, allowing researchers...high-resolution mapping of the brain by uniquely labeling neurons. Zebrafish are also amenable to manipulation of...consequences of activating or silencing specific neurons in living larvae. In addition, zebrafish have been...PubMed PMID: 18845710. Kim, Dal Hyung, et al. "Pan-neuronal calcium imaging with cellular resolution in freely...

...expressed may be found at very different levels (Mauro and Chappell, 2014). Due to this variation in tRNA...scientists with the ability to selectively silence neurons in vivo (Slimko and Lester, 2003).  Addgene has...modification sites and even affect protein function (Mauro and Chappell, 2014). Different rates of translation...experiments and in biotechnology and therapeutics (Mauro and Chappell 2014).  Ultimately, it is important...s12859-017-1793-7 Dresios J, Chappell SA, Zhou W, Mauro VP (2005) An mRNA-rRNA base-pairing mechanism for...604–615 . https://doi.org/10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon...dramatically improves expression levels. Journal of Neuroscience Methods 124:75–81 . https://doi.org/10.1016/...

...spinal muscular atrophy (SMA)." European Journal of Paediatric Neurology 19 (2015): S64-S65. Additional...results in the degeneration of spinal cord motor neurons. This disease is estimated to affect anywhere from...screens for small molecules that block the neurodegenerative effects of smn1 disruption. As part of his...for use in the clinic and thereby restore motor neuron function in spinal muscular atrophy patients. Noushin... University of British Columbia Department of Urological Sciences. Dr. Nabavi studies peritoneal mesothelioma...

...mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1-expressing mouse neuron without...optogenetic silencer in experiments with mouse cortical neurons, far out-performing previous attempts to engineer...of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential frequencies with ...channels that mediate optogenetic inhibition. Nat Neurosci 25, 967–974 (2022). https://doi.org/10.1038/s41593... of Addgene's more than 50 antibodies target neuroscience-related proteins, but we recently added several...

...of Experimental Methods. Neuron 90(3):434-6. https://doi.org/10.1016/j.neuron.2016.04.030    Bradbury ... to the catalog. The Catalog Will Start With Neuroscience Antibodies and Grow From There Addgene’s antibody...Brain Research Through Advancing Innovative Neurotechnologies® (BRAIN) Initiative. Our first collection ...materials is being developed for scientists studying neuroscience and is available to both academic and industry...Addgene will continue to add more antibodies, for neuroscience research and other disciplines, over time. The...

...findings relevant to my background (neuroimaging, biomechanics, neuropsychology) and from areas in which I knew...sentiment). Entering the final semester of a neuroscience major, I was struck with the realisation that...that I didn’t want a career in neuroscience—whatever that entails. Despite being engrossed by the theory...spheres of research. In my case, an interest in neuroscience was carried over and allowed to percolate within... notion of “heading” in soccer as a possible neurological health risk. Having played the sport my entire...

... split botulinum neurotoxin halves to enable VAMP2 cleavage, and suppresses neuronal transmission within...core of this tool is the endoprotease Botulinum Neurotoxin B (BoNT/B), which cleaves the synaptic protein...VAMP2 renders it inactive, thus inhibiting neurotransmitter release. How is this light-mediated? The researchers...photodimerizer system CRY2-CIB1 to regulate botulinum neurotoxin were also created. The depositors demonstrated...split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter...

...second vector that delivers gRNA, such as lentiGuide-Puro. The alternative single-vector systems typically...signaling pathways in specific cell types (typically neurons). While all of these viruses are popular, pAAV-...pAAV-hSyn-DIO-hM4D(Gi)-mCherry can be used to induce neuronal silencing in vitro and in vivo (Krashes et al..... The human synapsin promoter further specifies neuron-specific expression of the DREADD (Kügler et al...Lowell BB. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Invest..... Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an...

... RNA-sequencing to identify neuronal class and deep sequencing of neurons from each class to fully characterize...Addgene.  C. elegans open resources The C. elegans Neuronal Gene Expression Map & Network (CeNGEN) The CeNGEN...comprehensive gene expression atlas of every C.elegans  neuron. Scientists are currently doing this using two ...to view the anatomy of worm layer by layer, the NeuroML C. elegans connectome, a model of the C.elegans...Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed PMID: 30092212. PubMed...

...stimulation is a powerful tool used by neuroscientists to interrogate neuronal networks. A major drawback of this... method is the stray excitation of neighboring neurons in opsin dense regions. To overcome this issue,... C-terminus of the protein. Unlike the diffuse neuronal expression observed with wild type CoChR, soCoChR...describe a novel system for the precise mapping of neuronal networks at the single-cell level. Various soCoChR...available at Addgene. Shemesh OA, et. al. Nature Neuroscience. 2017. PubMed PMID: 29184208.        Synthetic...