...polynucleotide kinase 165521 pOpen-lambda red operon lambda red operon 165530 pOpen-T4-BGT T4-BGT beta-glucosyltransferase...

...of the DNA-binding domain of a TALE, along with strong nuclear localisation tags and repress transcription...transfected cells. TALEN expression is driven by the strong CAG promoter or can be achieved by in vitro mRNA...

...CRISPR/Cas Toolkit Kanemaki Lab Auxin-Inducible Degron Tagging Masato Kanemaki's lab has developed a simple...homology arms. Notably, they have used Auxin-inducible degron (AID) tagging with CRISPR/Cas to generate conditional...

...Gateway recombination-based cloning of complex polycistronic lentiviruses. Can express Cas9 with up to 3 ...in a single vector. sgRNAs are expressed as polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG ...

... related to cell-to-cell communications and environmental sensing. Sensing and Signaling Plasmids Search...

...transcription factors Connie Cepko See Plasmids Dronpa Optical control of protein activity Michael Lin...

...engineering: A new stem cell initiative from the NIH. Neuron . 2021 Apr 7;109(7):1080-1083. PMID: 33831364 (...

...treatment of neuroendocrine cancers. As such, NETRF strongly encourages its funded scientists to make their...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...homology-directed repair (HDR). HDR is much less error-prone than NHEJ and can be used to faithfully introduce...

... almost unidirectional insertions, but are more prone to off-target insertions. CASTs are most often used...

...blocks the progression of the RNA polymerase, thus strongly decreasing the expression level. Mismatches between...

...Crosnier C, Bustamante LY, Bartholdson SJ, Bei AK, Theron M, Uchikawa M, Mboup S, Ndir O, Kwiatkowski DP,...

...group A, member 1 PGR Plasmids PGR, NR3C3, PR progesterone receptor PPARA Plasmids PPARA, SC22CB-5E3.5,...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...Acknowledgement (RSA). This form can be completed electronically by the scientist who will be using the materials...

...Use our chemogenetics AAV to chemically induce neuronal activity in specific cell types. See our Chemogenetics...

...Choi J, Jones S, Ghebranious N, Igelmann H, Lu X, Soron G, Cooper B, Brayton C, Park SH, Thompson T, Karsenty...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...new technology, and we hope to continue to have a strong scientific partnership with both the labs that ...

...given donor-acceptor pair, the FRET efficiency strongly depends on the distance between the donor and ...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking...

...buffers. All buffers are supplemented with 0.001% pluronic F68 or 0.014% Tween 20. Phosphate Buffered Saline...

...construct ; pMA122 (negative selection marker); pGH8 (neuronal co-injection marker); pCFJ104 (body wall muscle...

...mutated allele may be different owing to the error-prone nature of NHEJ. In HDR gene editing experiments,...

...through non-homologous end joining (NHEJ) , an error-prone process that produces insertions/deletions disrupting...

...Dammeyer Lactose/IPTG inducible vectors with LacIq, a stronger repressor than LacI that lowers basal protein ...

...these regions, a process driven by neurons. By studying how neurons are physically connected, we can begin...these connections in the brain are (drum roll) neurons! Neurons are really cool because they can transmit ...There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to ... functionally dissect specific neurons within their natural environment, all while reciting their favorite...defined population of neurons. For example, say I wanted to access all the neurons that signaled from the...ligands activates both neurons at the injection site and any projection neurons retrogradely transduced... efficient retrograde access to projection neurons." Neuron 92.2 (2016): 372-382. PubMed PMID: 27720486...

.... What do microbial rhodopsins and neurons have in common? Neuronal stimulations are triggered by the ...the neuron fires, sending the electrical signal down the axon (Lodish et al., 2000). Most neurons in the...Vandenberghe, 2019). How to achieve strong opsin expression in specific neuronal cell types then? This is where... 2014). How does FLEx enable strong opsin expression in specific neuronal cells? This can be achieved ...enabling its expression under a strong promoter only in specific neuronal cells. FLEx ensures, therefore...et al. "Silencing neurons: tools, applications, and experimental constraints." Neuron 95.3 (2017): 504-...What is optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University...

...specify and assemble diverse arrays of neurons, forming the neuronal circuits that will later be refined ...individual neuronal populations.  The Yang Lab’s AAV plasmids used to express Cre under neuron-specific ...kinase II alpha (aCaMKII; excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs...response to neuronal activity (hence, “immediate-early”) and consequently regulates neuronal physiology...between Bdnf I-overexpressing neurons and Bdnf IV-overexpressing neurons. Among the upregulated genes ...contributions of transcript isoforms to neuronal physiology. The Day Lab’s neuron-optimized lentiviral vectors can...process. However, while the neuronal marker TUJ1 is useful for visualizing neuronal differentiation by immunohistochemistry...

...CNS: neurons and astrocytes Mice No (LY6C1)   Hanlon et al., 2019 AAV-BI-28 CNS: neurons, astrocytes...astrocytes, but its neuron transduction encompasses more molecularly-defined neuronal subtypes and with ...and is highly specific for neurons, with less bias toward inhibitory neurons than AAV-PHP.eB (Fig. 4c, ...transduce 80% of dorsal root ganglia (DRG) neurons, as well as neurons located in the enteric nervous system...primate nervous systems. Neuron, 110(14), 2242-2257.e6. https://doi.org/10.1016/j.neuron.2022.05.003 Chen, X...-specific labeling. Neuron,109(9), 1449–1464.e13. https://doi.org/10.1016/j.neuron.2021.03.011 Hanlon,...cell type specificity, or retrograde transport in neurons. (b) Genetic cargo can be customized to control...

...individual neurons and in terms of their interactions with one another. In addition to the neuronal networks... (hM3Dq) into a subset of neurons within the LC of mice. When those neurons are chemogenetically activated...reward circuit, and the medial prefrontal cortex (mPFC) are involved. Neurons from the mPFC project to the...findings by controlling neurons in the mPFC with light. Activation of these neurons via the channel Channelrhodopsin...protein that activates neurons upon light stimulation. When they activated the Drd1 neurons 24 hours before ... "Optogenetic stimulation of medial prefrontal cortex Drd1 neurons produces rapid and long-lasting antidepressant...They also don’t give any clues about the types of neurons involved or whether their involvement is activating...

...for genetically manipulating neurons throughout the whole brain. Neurons of the peripheral nervous system...transduced a higher number of neurons overall (GFP+ cells that also stained for a neuronal marker). Check out tables...for?  1. Multi-color labeling of neurons Multi-color labeling of neurons is for more than making pretty ...single-cell anatomical study and neuronal tracing, aka determining where a neuron’s axons and dendrites extend...in order to decrease the number of neurons that get labeled, neurons are more likely to be transduced by...interest, which, in the case of Chan et al, was neurons and astrocytes. Why screen for IV deliverable ...and GFP intensity and transduction efficiency of neurons were measured. On average, AAV.PHP.eB and AAV.PHP.S...

...method that lets you fire neurons with the flick of a light switch. Neurons are not typically persuaded...remains inactive and neurons don't fire. With blue light, ions flow into the neuron and causes it to fire...ChRger. In both experiments, neurons expressing ChRgers produced stronger photocurrents at low light intensity...activate a neuron. ChRgers, however, have a higher conductance, so they can still depolarize neurons at lower...dopaminergic neurons in the VTA, this should propel the mice to increase their nose-poking behavior. Strong nose-poking...shines on excitatory channelrhodopsin-expressing neurons, the channelrhodopsins respond by opening and allowing... allowing an influx of ions into the neuron which generates an action potential (Figure 1). This light-induced...

...periaqueductal gray (PAG), with red neurons expressing tdTomato and green neurons expressing cFos. Image from ...the activity of parvalbumin (PV)-expressing neurons, neurons that the lab previously found had a role in...light activates the neurons (via oChIEF) and a chemogenetic agonist inhibits the neuron (via hM4D). She fused...on the PV neurons is in progress, she has validated the dual virus method to study CR neurons which are...fluorescence based on the release of glutamate from neurons and this new version of the biosensor is faster... to the collaborative and highly interactive environment that Janelia offers,” says Aggarwal.In addition... in electroporated in rat hippocampal cultured neurons, imaged 14 days post-transfection, excited at 505...

...Genetically Targeted Neurons. Neuron 53:639–647 . https://doi.org/10.1016/j.neuron.2007.01.033 Additional...the previous neuron (Wichersham et al., 2007). However, once rabies dG enters this neuron, it cannot infect...However, pseudotyping allows these viruses to infect neurons. Besides altering the host tropism of your virus...reasons VSV-G is used to transduce stem cells and neurons. However, VSV-G’s broad tropism can be undesired...Rabies glycoprotein The rabies glycoprotein targets neuronal cells and can travel in a retrograde direction...the cell body, making it a great tool to study neuronal connectivity. However, because of rabies virus...Lentiviral vectors are also great at infecting neurons and cause minimal inflammatory response, but are...

...they make the inside of the neuron more positive or more negative.  Neuronal stimulation: Depolarization...depolarization. They therefore function as a neuron “on switch.” Neuronal inhibition: Hyperpolarization Conversely...cord neurons using the Gal4/UAS system. This created fish lines with different groups of neurons that ...activity, enables researchers to silence or incite neuronal firing and study subsequent effects on behavior...cross the cell membrane when exposed to light. Neuronal firing depends on these charged particles and ...Depolarization When the inside of the neuron becomes positive over a certain threshold, it becomes depolarized and...Conversely, when the inside of the neuron becomes more negative it becomes less likely to fire and is said...

...reversible PA-FP is the naturally occurring Dronpa protein (named after dron, a ninja term for vanishing, and pa...both activates and inactivates Dronpa is not ideal, since this means Dronpa is quickly inactivated and less... to find a version of Dronpa that doesn't become inactivated as quickly. Padron is a monomeric, reversible... mutating Dronpa (T59M / V60A / N94I / P141L / G155S / V157G / M159Y / F190S) (7). Padron’s “positive”...switching behavior is the opposite of Dronpa; when exposed to blue light, Padron is photoconverted from a dark...Emission (nm) Dronpa Monomer UV-Violet Dark/Green N/A (high) High 503/ 518 Padron Monomer Blue Dark...Correlative light and electron microscopy (CLEM) is an imaging technique that combines electron microscopy (EM...