Neuroscience Research Applications for Gene Synthesis

Overview

Gene synthesis is widely used in neuroscience research studies: codon-optimized channelrhodopsins enable optogenetic approaches to visualizing neuronal pathways; customized ORF clones and RNAi constructs elucidate glioblastoma signaling pathways; and gene variants are used to investigate neuronal signalling mechanisms that underlie synapse plasticity and learning behaviors, just to name a few. GenScript is the most cited gene synthesis provider in the world, with over 300 neuroscience publications citing our gene synthesis services and other bio-reagent services and products. Read more to see why neurobiologists around the world use gene synthesis to help accelerate their research.

Explore recent publications that used gene synthesis, or read about featured research news topics below.

• Chuong et al. Noninvasive optical inhibition with a red-shifted microbial rhodopsin. Nat Neurosci. 2014 Aug;17(8):1123-9. Read full text
• Zimmermann et al. Synaptobrevin 1 mediates vesicle priming and evoked release in a subpopulation of hippocampal neurons. J Neurophysiol. 2014 Jun 18. Read full text
• Watt et al. Do current therapeutic anti-Aβ antibodies for Alzheimer's disease engage the target? Acta Neuropathol. 2014 Jun; 127 (6); 803-10.  Read full text
• Hansen et al. Distinct functional and pharmacological properties of Triheteromeric GluN1/GluN2A/GluN2B NMDA receptors. Neuron. 2014 Mar 5;81(5):1084-96. Read full text

Alternative Splicing Boosts Brain Size

mammalian-specific splice variant of PTBP1 drives neurogenesis

Humans have about the same number of genes as simpler vertebrates – so how does our genome give rise to such a stunning diversity of proteins, brain structures, and ultimately to the behaviors that make us human? Alternative splicing plays a big role, as a new paper published in Science shows – and a single exon-skipping event in a critical regulatory protein PTBP1, drives widespread changes in splicing that could account for the huge differences in brain development across vertebrate species.

By expressing a mammalian-specific splice variant of PTBP1 in chickens, scientists gained clues into why mammals develop larger brains than other vertebrates. PTBP1 regulates the splicing of hundreds of downstream target genes, and the mammalian-specific splice variant activates a brain-specific program of alternative splicing, favoring the creation of new neurons. Further studies will investigate how the different forms of PTBP1 hand off control during specific stages of embryonic development to regulate neurogenesis and brain size.

GenScript offers molecular biology services to accelerate research on alternative splicing and the molecular mechanisms governing neurogenesis and brain tissue patterning:

• Gene synthesis, with industry-leading codon optimization, starting at $0.23/bp
• Express Cloning: 151 free vectors to choose from to get expression-ready clones for just $49/clone
• CRISPRgRNA constructs: validated gRNA sequences in your choice of vector for $199/clone

Optogenetics Goes Two-Channel

New opsins Chronos and Chrimson solve the problem of how to excite distinct neuronal populations.

"Optogenetics" allow researchers to stimulate synaptic activity in specific neurons that are made to express light-sensitive ion channels (channelrhodopsins). Despite the variety of opsins with different peak wavelength sensitivities, until now it has not been possible to independently activate two distinct neural populations without significant cross-talk or losing temporal resolution. Researchers at MIT reported in Nature on two new opsins with non-overlapping excitation spectra, Chronos and Chrimson, that allow independent optical excitation of distinct neural populations in mouse brain slices. These tools open the door to explore how multiple synaptic pathways interact to encode information in the brain.

Opsin genes occur naturally in microbial algae. In order to efficiently express these genes in mammalian cells, these researchers turned to GenScript for codon optimization and gene synthesis. GenScript has developed the leading codon-optimization algorithm: OptimumGene, patented in 2012 and continuously improved based upon the latest research findings.

• Read the full publication: Klapoetke et al. Independent optical excitation of distinct neural populations. Nat Methods. 2014 Mar;11(3):338-46
• See more articles on optogenetics
• Request a free codon optimization report for your gene sequence.

A Newly Identified Post-Translational Modification Critical for Learning

δ-catenin palmitoylation mediates activity-induced synapse plasticity

Synaptic plasticity is known to underlie learning and memory, but we're still discovering the molecular mechanisms by which synaptic activity leads to changes in synapse morphology and function. A recent Nature Neuroscience paper reports a new post-translational modification that is required to coordinate the changes involved in memory formation: after enhanced synaptic activity, DHHC5 palmitoylates δ-catenin, increasing its binding to synaptic cadherin both in vitro and in the hippocampus of fear-conditioned rodents. These researchers used a synthetic δ-catenin gene, in which the palmitoylated cysteine residues were mutated to serine, to demonstrate that δ-catenin palmitoylation is required for numerous facets of activity-induced synaptic plasticity, including N-cadherin stabilization at synapses, postsynaptic spine enlargement, insertion of GluA1 and GluA2 into the synaptic membrane, and increased mEPSC amplitude.

• Gene Synthesis is the most efficient way to generate the DNA constructs your research requires.
• See over 300 neuroscience papers that cite GenScript

A New Candidate Drug Fights Glioblastoma by Targeting a Suite of miRNAs that Regulate Wnt/β-catenin Signaling

The small molecule BASI showed potent anti-cancer effects in a high-throughput screen for glioblastoma multiforme (GBM), the most common type of adult brain tumor. Now a new study reveals that BASI inhibits glioma proliferation, invasion, and migration in vitro and in vivo by altering levels of several micro-RNAs, which form a coordinated, multi-level blockade of the Wnt / β-catenin signaling pathway. Building upon prior work that elucidated the role of several of these miRs in regulating Wnt signaling, this study provided novel characterization of the role of miR-181d, and reported on the ability of the small molecule BASI to simultaneously target the entire suite of miRNAs that regulate β-catenin through diverse mechanisms. Because Wnt is implicated in the growth of multiple tumor types, BASI and the miRs it regulates are promising subjects for further basic and translational research.

• miR-181d suppresses glioblastoma progression by directly targeting CTNNB1 (the gene that encodes β-catenin)  and CREBBP (the gene that encodes CBP).
• miR-200a directly interacts with the 3'-UTR of CTNNB1 to suppress gene expression.
• miR-21 directly targets von Hippel-Lindau (VHL) and peroxisome-proliferator-activated receptor α (PPARα), thereby regulating VHL-mediated β-catenin degradation as well as EGFR/AKT signaling through VHL/β-catenin and the PPARα/AP-1 axis.^Ref
• inhibition of miR-23b reduces tumor malignancy by downregulating HIF-1α, β-catenin, MMP2, MMP9, VEGF and ZEB1 and increasing expression of VHL and E-cadherin.^Ref

To study the effect of miR-181d on the β-catenin pathway, these researchers ordered custom-cloned CTNNB1 and CREBBP reporter plasmids from GenScript. With CloneEZ™ Molecular Cloning, it's easy to get customized ORF clones or even completely custom genes in expression-ready plasmids.

• Read the full text of the paper.
• GenScript has CTNNB1 ORFs in stock, plus ORF clones for 151 genes in the WNT signaling pathway. To get ORFs cloned into your choice of vector starting at $199 and 10 day-turnaround time, try our CloneEZ™ ORF cloning service.

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