...in the puromycin cassette that makes it less efficient in some cell lines. pSpCas9(BB)-2A-Puro (PX459)...Find pLKO.1 - TRC cloning vector. Tet-pLKO-puro - Tet-pLKO-puro contains all the necessary elements for ... by the Weiderschain lab. Find Tet-pLKO-puro. lentiGuide-Puro - This plasmid expresses the S. pyogenes.... Find lentiGuide-Puro. pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)-2A-Puro (PX458), pSpCas9n...plasmid pSpCas9(BB)-2A-Puro (PX459) V2.0. It contains a point mutation in the puromycin cassette that makes...best to go with pSpCas9(BB)-2A-Puro (PX459) V2.0. Find pSpCas9(BB)-2A-Puro (PX459). Thank you! We would...pX330-U6-Chimeric_BB-CBh-hSpCas9. pSpCas9(BB)-2A-Puro (PX459) V2.0 - This plasmid from the Zhang lab is...

...within a neuron? For my PhD I studied a synaptic protein that is linked to neurodegeneration. The level... Rojas, PhD is a neuroscientist who has always been deeply interested in neurobiology.  She is passionate...synthesized proteins in rat hippocampal neurons." Nature neuroscience 13.7 (2010): 897-905. PubMed PMID: 20543841...Nathans, Daniel. "Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains... to study how protein turnover is regulated in neurons. Protein turnover is the combined effect of protein... I wanted to measure it’s levels in its normal neuronal context. In other words, I needed a method for...for visualizing protein turnover in situ in neurons. The classic approach for studying protein turnover...

...2011) labels Drosophila neurons at the neuroblast stage, meaning that neurons of the same lineage will...multicolor labeling is a popular technique in neuroscience and developmental biology. This type of cell...cell types, and research applications. Labeling neurons with Brainbow The Lichtman group developed the ...iteration of this technology in 2007. Used to label neurons in the cerebellum of mice, the team appropriately...2007). Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent proteins are organized...the truncated YFP. In Thy1-Brainbow mice, some neurons expressed as many as 16 copies of the injected ...multicolor labeling of the brain. Labeling Drosophila neurons with dBrainbow and Flybow Based on the Brainbow...

...proteins, work in different neurons and neuroglia, and have different effects on neuron activity (firing or inhibition...2016) DREADDs for Neuroscientists. Neuron 89:683–694 . https://doi.org/10.1016/j.neuron.2016.01.040 Takeda...in a specific neuronal subtype in the brain that can either activate or inhibit neurons. You can find ...initiating signaling pathways in inflammation, and neurotransmission. These receptors can be activated by an array...of different ligands, including hormones, neurotransmitters, photons or odorants, to elicit downstream...chemogenetic studies to probe the relationship between neurons and specific behaviors. Chemogenetics uses genetically... small molecules to either activate or inhibit neuronal firing. As GPCRs are the largest class of signal...

...to monitor? Neurons, for example, have unusually fast calcium dynamics, whereas non-neuronal systems tend...populations of neurons in vivo to study circuit activity. CaMPARI is available for neuron-specific expression...in molecular neuroscience. 2014 Nov 18;7:88. Tian L, Hires SA, Looger LL. Imaging neuronal activity with...and target a defined population of cells (e.g., a neuron subtype) and specific subcellular compartments ... rapid measurements, like action potentials in neurons. GCaMP7 and GCaMP8 are also available with improved... studies. A particularly useful sensor for neuroscientists is CaMPARI. This sensor switches from green...improved version named CaMPARI2 is available for neuron-specific expression in both zebrafish and mammalian...

...in MeCP2 null (S51.002). Neurology 92 (15 Supplement) . http://n.neurology.org/content/92/15_Supplement...gene, Rett Syndrome is a rare, progressive, neurodevelopmental genetic disorder that affects primarily girls...CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson & Bird, 2019) and is a global...silenced by a Lox-Stop cassette, resulting in the neurological deficits observed in Rett Syndrome. Restoring... tamoxifen-inducible Cre not only halted the neurological damage but completely reversed it, regardless...the MECP2 protein is necessary to rescue the neurological defects of Rett syndrome (Tillotson et al., ...results for repairing mutations in models of neurological disease. Klaus Rajewsky’s group recently used...

... of densely labeled neurons because background fluorescence from inactive neurons is reduced. 105322... of densely-labeled neurons because background fluorescence from inactive neurons is reduced.  Additional...calcium sensors and can be used for imaging of small neuronal processes like dendrites and axons. Find the 7b... sensor AAV GABA is the primary inhibitory neurotransmitter in vertebrates, but methods for directly imaging...Archon1 voltage reporter allows a way to monitor neuronal electrical activity through imaging. Since it’...mDlx enhancer restricts expression to GABAergic interneurons. These vectors can also be used as the control...flex PSAM4 GlyR IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR...

... of densely-labeled neurons because background fluorescence from inactive neurons is reduced. This variant...inhibitory neurotransmitter - is a powerful method for understanding how this molecule impacts neuronal circuits...50457-AAV5): The Synapsin promoter directs broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5...element directs expression in forebrain GABAergic interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP...(jGCaMP7) continue to be useful for monitoring neuronal activity in the form of calcium dynamics. (For...fluorescence and can be used for imaging small neuronal processes (dendrites and axons). jGCaMP7c exhibits...to be a useful method for manipulating specific neurons. We recently packaged a soma-targeted opsin from...

...-IRES-Puro: Expression of the SARS-CoV-2 N protein  pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro: Expression...pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV-2-nsp2-2xStrep-IRES-Puro) are also in the works...continues to supply neuroscience with ever more efficient tools for light mediated neuronal control. The advantages...protein pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a Lentiviral ...plasmid for expression in forebrain GABA-ergic interneurons (83895). See our Serotype Testing AAV inventory...system that allows for specific manipulation of neurons. We have expanded our AAV collection to include...targets expression of ChR2 to forebrain GAB-ergic interneurons through the mDlx enhancer element. Plasmid...

...integrating the GFP-Puro cassette into the FBL locus. Upon sequencing of puromycin resistant clones, they...GFP-Puro will be inserted just upstream of a stop codon. One potential concern is if the GFP-Puro will...detailed protocol for MMEJ-mediated knock-in of a GFP-Puro cassette into a given locus, just upstream of a ...microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks are necessary...necessary for knock-in: one on either side of the GFP-Puro cassette and one in between the 5’ and 3’ microhomologies...microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the locus (see figure below). The ...bp 5’ and 3’ microhomologies are added to the GFP-Puro cassette via PCR, and this construct is inserted...

... Red Light Optogenetics in Neuroscience. Frontiers in Cellular Neuroscience, 15, 778900. https://doi.org...the first light-activated tool for controlling neuronal activity, channelrhodopsin-2 (ChR2), a blue light-activated... exposed to blue light, this channel activates neuronal activity by opening and allowing positively charged... action potential - a very useful tool for neuroscientists! Since then, scientists have developed many...channels (rhodopsins) either excite or inhibit neuronal activity upon exposure to light within a specific... when you want to activate or inhibit specific neurons without needing to consider secondary signaling...Optogenetics Resources and the Boyden lab’s Synthetic Neurobiology Group Resources, both of which have extensive...

...expression specifically in neurons. AAV Delivered Cre - A useful tool for neuroscientists AAVs are widely used...used by neuroscientists because they efficiently deliver genes to various brain regions. You can partially...way. For example, say you would like to label neurons with mCherry and you already have a transgenic ...entirely new mouse line with mCherry controlled by a neuron specific promoter or you could breed your Cre-dependent...line with a separate line that expresses Cre in neurons, but both of these options would be very time consuming...and therefore mCherry will only be expressed in neurons. Indeed, this AAV provides you with a means to ...expression of any Cre-dependent gene of interest in neurons. This ability to be paired with any Cre-controlled...

...improve the study of specific neuronal cell types involved in neurodegenerative diseases, the Cole Lab selected...linked to disease, including motor neurons (-3mnx1/Hb9), pan neuronal (elavl3/HuC), microglial/macrophage...specific neuronal subtypes was not available until now. In order to target specific neuronal subtypes ...infecting GABA-ergic interneurons here. Dimidschstein J, et al. Nature Neuroscience. 2016. PubMed PMID:...generate zebrafish transgenic lines to study neurobiology and neuorodgenerative diseases. The Tol2 Gateway-Compatible...), and astrocytic (gfap).This toolbox adds new neuronal tools to the expanding gateway compatible vectors... PMID: 25772473 Targeting and manipulating interneurons using a viral approach Genetically modified...

...great in a GABA neuron, for instance, may not transduce the adjacent norepinephrine neuron at all. There... for Defense Behaviors. Neuron, 93(1), 33–47. https://doi.org/10.1016/j.neuron.2016.11.045 More resources...expression can still occur (Fig. 3) - for example, some neurons can transiently express TH - so you’ll need to ...expression in both oligodendrocytes (Olig2) and neurons (NeuN). Image from Griffin et al., 2019, Figure...central nervous system. Frontiers in Molecular Neuroscience, 7. https://doi.org/10.3389/fnmol.2014.00076...cFork anterograde tracing system. Experimental Neurobiology, 29(3), 219–229. https://doi.org/10.5607/en20006...

...named NeuroStack was used (Orguc et al., 2020). Using a programmable microcontroller, NeuroStack coordinates...and stimulate or manipulate responses through neurological means like electric stimulation, drug administration...optogenetic tools, calcium sensors, and other common neuroscience tools into the brains of model organisms, including...tissue. They performed optical stimulation using NeuroStack and the fibers as previously described, then ...trended with expression of the opsin in the target neurons, validating the functionality of these inputs and...Success! They also observed reliable optically evoked neuron activity for at least several months after implantation...enteroendocrine cells within the intestines known as neuropods. They observed increased vagus nerve firing in...

...biologists, while neuroscientists would like to monitor calcium specifically in their favorite neurons. This specificity... V., Fluorescent Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers... or animal system. This ‘thing’ could be a neurotransmitter, such as norepinephrine, or an electrolyte...contraction, and calcium channels are activated for neuronal action potentials; it’s an ion of interest for...—keep reading to find out how!   Neurotransmitters Many canonical fluorescent as well as FRET-based...been engineered to detect a wide array of neurotransmitters ranging from dopamine to glutamate, all the... Frontiers in Cellular Neuroscience. 2019 Oct Vol. 13; 1662-5102. DOI: 10.3389/fncel.2019.00474 Resources...

...cell types (astrocytes, microglia, neurons, etc.) and even neuronal phenotype in a mixed population (Burger...Many neuroscience experiments that require gene expression in a specific cell type rely on transgenic...tyrosine hydroxylase (TH) promoter for catecholamine neurons or glial fibrillary acidic protein (GFAP) for astrocytes...vectors for off-target expression.  Some classes of neurons transiently express TH during development or have...there is promoter activity. Additionally, some neurons express low levels of GFAP, which can lead to off-target...off-target expression of the packaged transgene in neurons and glia.   Check the recombination rate The first...on Helper Virus Concentrations. Front Synaptic Neurosci 12. https://doi.org/10.3389/fnsyn.2020.00006 O...

...oligodendrocytes, microglia, astrocytes, neurons, and inhibitory neurons. Your choice of AAV will depend upon... as a new tool for neuroanatomical tracing (11). With the ability to produce neuron specific transfection...contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University of Manitoba. ... These AAVs may move toward the cell bodies of neurons (retrograde transport) or towards the synapse (...Cre-lox, this new method could allow for labeling of neurons showing a very specific phenotype. Previous neural... Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student at the University of Manitoba. ... Floris G., et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring...

...coffee does it take to fuel more than 30,000 neuroscientists over a 5 day conference? I feel like here at... at the annual Society for Neuroscience meeting in Washington, D.C., we must be making a dent in the global...conference, I have been impressed by how the neuroscience community pulls off such an inspiring, thought-provoking...made in reprogramming astrocytes to functional neurons in vivo. And I’ve seen evidence of the infiltration...infiltration of CRISPR-Cas tools into neuroscience experiments. Speaking of CRISPRs, we were excited to have... the importance of scientific sharing to the neuroscience community. The scientists we've spoken with ... for a free plasmid! Also check out the PLOS Neuroscience Community blog to read reports from bloggers...

... 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...