...pouring in! While our lentiviral service is going strong, the AAV service has shown incredible growth this...be used to drive Cre expression specifically in neurons. AAV Delivered Cre - A useful tool for neuroscientists...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...

...their environmental impact on a scale of 1-10: the lower the score, the lower the environmental impact...receive the product? How can we consider the environmental impact of those processes? Who even regulates...years since the creation of the United States Environmental Protection Agency, which started making regulations... can choose products that are better for the environment without sacrificing performance or quality. To...destined for a landfill. However, Triumvirate Environmental developed Red2Green, a program that uses this...lumber for things like truck bumpers, plastic adirondack chairs, and synthetic turf. I was truly amazed... a practice which we now realize is a major environmental disaster. We should continue to examine our ...

... make enzymes controllable by tethering them to Dronpa, a fluorescent protein known to oligomerize when...To do so, she generated fusion proteins in which Dronpa domains flanked either side of an enzyme’s active...ultraviolet light exposure caused oligomerization of the Dronpa domains thereby occluding the enzyme active site...the enzyme. Later exposure to cyan light stopped Dronpa oligomerization and turned the enzymes back on....optimized these first generation FLiPs by mutating Dronpa to lower its aggregation tendency, increase its.... She additionally showed that these enhanced “pdDronpa” domains could be placed in amino acid loops on...iterations of FLiPs only used N- and C-terminal dronpa fusions. With these various designs, Xin has been...

...Manon Eckhardt and Melanie Brewer from the QBI Coronavirus Research Group at UCSF. It’s been only a few ...-- all while maintaining social distance.  From front to back: David Gordon, Jeff Guo, Fengbo Zhou, Paddy...scientists from more than 20 labs to form the QBI Coronavirus Research Group (QCRG) to do just this.  In under... York. We want YOU to join the fight against coronavirus We are just at the beginning of understanding...stick together (6 feet apart), to fight the new Coronavirus. To help get you started, here are a few key ...clone selection. Expression is driven by a fairly strong EF1alpha promoter. We have verified expression ...lab's Addgene page to get notified!) Attacking coronavirus, one FedEx package at a time Since the publication...

...proteins, work in different neurons and neuroglia, and have different effects on neuron activity (firing or inhibition...in a specific neuronal subtype in the brain that can either activate or inhibit neurons. You can find ...DREADDs for Neuroscientists. Neuron 89:683–694 . https://doi.org/10.1016/j.neuron.2016.01.040 Takeda S, Kadowaki...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...concise table of DREADDS and their activity in neurons on Addgene’s chemogenetics guide.     References...

... Jason Moffat at the University of Toronto has created the Toronto KnockOut (TKO) CRISPR Library. This...for neuronal systems. To develop a new single-component tool for magnetogenetic control of neurons, the...sequences outside of their endogenous chromatin environment and relies on the fact that enhancers function...J Genet Genomics. 2016. PubMed PMID: 26887506 Toronto KnockOut (TKO) CRISPR library In order to identify...Natl Acad Sci USA. 2016. PubMed PMID: 26884157 Aronshtam, et al. Nucleic Acids Res. 1996. PubMed PMID: ...but turns more red in lower (e.g. vacuolar) pH environments that inactivate SEP and leave only DsRed to ...the Co-IP compared of a control Co-IP indicates stronger interaction. The system has been demonstrated ...

...inducible degron (AID) system in C. elegans By Alyssa Cecchetelli The auxin inducible degron (AID) system...Tracking cell-cell interactions New auxin inducible degron system for C. elegans New CRISPR plasmids New items...cell types including endothelial cell-pericyte, neuron-astrocyte, and cancer-stromal cell pairs.   ...   Schematic showing the auxin-inducible degron system. Image from Ashley et al., 2021.   The...spatiotemporal expression pattern and an auxin inducible degron (AID) sequence attached to your protein of interest...presence of auxin, this TIR1 complex binds to the degron sequence ultimately leading to the degradation ...the AID* (minimal 44 amino acids auxin inducible degron sequence) tagged proteins. Degradation of your ...

...activity of neurons on or off using pulses of light. By activating a neuron or set of neurons, you can see...activate a set of neurons, maybe we could treat patients suffering from Parkinson's, chronic pain, deafness...independently control the activity of two populations of neurons at once. Addgene spoke to Boyden about this latest...contributes to behavior or pathology, and by disabling neurons you can figure out what they are necessary for....examples where you want to activate two sets of neurons and see how different parts of the brain or the...

...Much later, it was shown that putting the operon under a strong promoter results in high R-body expression...machines that can extend and retract over many microns in response to pH changes without the need for ... are composed of 4 proteins encoded in a single operon and were first expressed in E. coli in the 1980s... scaffold that changes shape in response to environmental conditions. More speculatively, R bodies might...of armor to protect their host from predation (Petroni et al 2000). With a better understanding of how...new ways to interact mechanically with their environment.   Jessica Polka is a Postdoctoral Research ... Plasmids from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer, K.-H., Verni, F. & ...

...from a state of chronic hepatitis caused by hypernutrition and alcohol consumption. Chronic inflammation ...cancer and defeat it we need to attack it from all fronts!”  Here I aim to compile a few highlights from ...researchers are preparing and carrying out this multi-pronged attack. Inflammation and tumorigenesis The conference...about alcohol and obesity as driving factors for chronic inflammation which in turn suppresses cancer immunosurveillance...surface receptor PD-1. This immunosuppressive environment in turn may represent a favorable state for tumor...Resources. Studying the immune system and tumor microenvironment Tumor Immunosurveillance and Defense With ...Friedmann-Morvinski about tools used to analyse the tumor microenvironment (one of the plasmids on her poster is pTomo-Ras...

...our observations at the electron microscope showed us that, in monkey neurons, the large majority of the...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)...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... of interest the membrane.   Figure 1: Electron microscope images showing ultrastructural localization...protein of interest, an alternative could be bicistronic expression of the fluorescent protein with the...

...oligodendrocytes, microglia, astrocytes, neurons, and inhibitory neurons. Your choice of AAV will depend upon... thanks to our guest blogger Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student at ...post was contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University... These AAVs may move toward the cell bodies of neurons (retrograde transport) or towards the synapse (...a larger amount of vector using a serotype with strong CNS tropism (4). If axonal transport is not desirable...neuroanatomical tracing (11). With the ability to produce neuron specific transfection using Cre-lox, this new method... method could allow for labeling of neurons showing a very specific phenotype. Previous neural tracing...

...plasmids: Luc (intronless; CMV-LUC2CP/ARE and Luc-I (intron-containing; CMV-LUC2CP/intron/ARE). The Luc-I...Luc-I intron is a chimeric β-globin/immunoglobulin intron that has been commonly used in constitutive splicing...Multimerization via the SunTag system also shows strong upregulation of gene activation when fused to dCas9... simplifies the process of creating complex polycistronic lentiviruses. The MuLE toolbox contains a series...induced-pluripotent stem cells (iPSCs) to take on a “neuron-like” phenotype when targeting genes involved in...in mammalian cells, yeast, and Drosophila. Synchronization of secretory protein traffic in mammalian cells...disrupts this interaction and thus triggers a synchronous release of the reporters which can then be tracked...

...electrochemical sensors for emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067...variety of applications, including detection of environmental pollutants, drug discovery, and disease diagnosis...instead of protein-based fluorophores, including: Environments with higher temperatures, since proteins can...Jonathan. “RNA Fluorescence with Light-Up Aptamers.” Front Chem. 2016 Jun 28;4:29. PubMed PMID: 27446908. PubMed...systems using immobilized aptamers.” Biosens Bioelectron. 2011 May 15;26(9):3725-36. PubMed PMID: 21419619.... PubMed Central PMCID: PMC4800036. 12. Wang, Rongsheng E., et al. “Aptamer-Based Fluorescent Biosensors...assay for prion diseases diagnostic”. Procedia Environmental Sciences. Volume 12, Part B, 2012, Pages 1348...

... pSC101 copy number is regulated by iterons. RepA binds to iterons and recruits host proteins to the ori...is initiated at the ori and may or may not be synchronized with the replication of the host cell's chromosomal...RepA binds to directly repeated sequences called iterons located near the ori and directly interacts with...check, high copy numbers can allow RepA to link iterons from two different plasmids, forming a pair of ...either of them (Park et al., 2001). The number of iterons encoded on a given plasmid influences how high ...host, as no additional components are necessary. Iterons are not the only tool to stringently regulate plasmid... numbers, RepA proteins form complexes between iterons on different plasmids in a process called "handcuffing...

...factor RepA binds to the iteron to initiation replication. However, in iteron-containing plasmids,  RepA... regulation system involving antisense RNAs or iterons that inhibits replication when copy number in the...   Figure 1: The replication problem. Iterons are repeated sequences within the origin of replication...del Solar et al., 1998). There are two models for iteron-based plasmid replication control that are not ...the second model, termed the “handcuff” model, iterons of two separate plasmids bind to the same RepA ...these plasmid types? Read our Plasmids 101: Environmental Plasmids blog). Scientists have designed small...replication and transfer systems and host taxonomy. Frontiers in Microbiology 6: . https://doi.org/10.3389/fmicb...

...tobacco. Studying gene function in C. roseus is strongly limited by the availability of methods. Infection...reporter gene examples are below. The GUS gene (β-glucuronidase) with histochemical GUS-staining is a very useful...days in the dark after the transformation. Use introns Plant promoters can be recognized or leaky in Agrobacterium...Agrobacterium, but not the plant. For example, we have seen strong expression of the GUS gene with the commonly used...Agrobacterium. This can be easily avoided by the use of introns, which will be removed from the transcripts in ...plants, check out this interview with the Nicola Patron, who shared the MoClo plant kit via Addgene. For...well working protocol in your lab, it can become a strong tool for your research. Feel free to add further...

...neural fibers developed by the Anikeeva lab are microelectronic fibers made from thermoplastic polymers that...desired thickness. The fibers are drawn with microelectronics included in them for downstream control – ...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..., P., & Chandrakasan, A. P. (2020). Modular Optoelectronic System for Wireless, Programmable Neuromodulation... Anikeeva, P. (2019). Flexible fiber-based optoelectronics for neural interfaces. Chemical Society Reviews...A., … Anikeeva, P. (2023). Multifunctional microelectronic fibers enable wireless modulation of gut and...

...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...channels (rhodopsins) either excite or inhibit neuronal activity upon exposure to light within a specific...hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation and inhibition by ... when you want to activate or inhibit specific neurons without needing to consider secondary signaling... off-target activity. Whether you are studying neuronal signaling or cancer pathways there are a number...2022). Red Light Optogenetics in Neuroscience. Frontiers in Cellular Neuroscience, 15, 778900. https://...

... How do you visualize protein turnover within a neuron? For my PhD I studied a synaptic protein that is... 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... whole population of one particular protein in neurons or in any other cell type. However, we cannot distinguish... newly synthesized proteins in rat hippocampal neurons." Nature neuroscience 13.7 (2010): 897-905. PubMed...Robert H. Singer. "Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability...