ArticleMolecular brain2015
The production of viral vectors designed to express large and difficult to express transgenes within neurons.
Article in Molecular brain, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 24 citations in OpenAlex.
- Advances in HIV Gene Therapy.International journal of molecular sciences · 2024Review
- Functional genome-wide short hairpin RNA library screening identifies key molecules for extracellular vesicle secretion from microglia.Cell reports · 2022Article
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- Regulatory Architecture of the Neuronal Cacng2/Tarpγ2 Gene Promoter: Multiple Repressive Domains, a Polymorphic Regulatory Short Tandem Repeat, and Bidirectional Organization with Co-regulated lncRNAs.Journal of molecular neuroscience : MN · 2019Article
- Therapeutic potential of combined viral transduction and CRISPR/Cas9 gene editing in treating neurodegenerative diseases.Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology · 2018Review
- Development of lentiviral vectors for efficient glutamatergic-selective gene expression in cultured hippocampal neurons.Scientific reports · 2018Article
- The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to NeuronsFrontiers in molecular neuroscience · 2018Article
- Genome modification of CXCR4 by Staphylococcus aureus Cas9 renders cells resistance to HIV-1 infection.Retrovirology · 2017Article
- A lentiviral vector bearing a reverse intron demonstrates superior expression of both proteins and microRNAs.RNA biology · 2017Article
- The Development of a Viral Mediated CRISPR/Cas9 System with Doxycycline Dependent gRNA Expression for Inducible In vitro and In vivo Genome Editing.Frontiers in molecular neuroscience · 2016Article
- Overexpression of NTRK1 Promotes Differentiation of Neural Stem Cells into Cholinergic Neurons.BioMed research international · 2015Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
backgroundViral vectors are frequently used to deliver and direct expression of transgenes in a spatially and temporally restricted manner within the nervous system of numerous model organisms. Despite the common use of viral vectors to direct ectopic expression of transgenes within the nervous system, creating high titer viral vectors that are capable of expressing very large transgenes or difficult to express transgenes imposes unique challenges. Here we describe the development of adeno-associated viruses (AAV) and lentiviruses designed to express the large and difficult to express GluN2A or GluN2B subunits of the N-methyl-D-aspartate receptor (NMDA) receptor, specifically within neurons.
resultsWe created a number of custom designed AAV and lentiviral vectors that were optimized for large transgenes, by minimizing DNA sequences that were not essential, utilizing short promoter sequences of 8 widely used promoters (RSV, EFS, TRE3G, 0.4αCaMKII, 1.3αCaMKII, 0.5Synapsin, 1.1Synapsin and CMV) and utilizing a very short (~75 bps) 3' untranslated sequence. Not surprisingly these promoters differed in their ability to express the GluN2 subunits, however surprisingly we found that the neuron specific synapsin and αCaMKII, promoters were incapable of conferring detectable expression of full length GluN2 subunits and detectable expression could only be achieved from these promoters if the transgene included an intron or if the GluN2 subunit transgenes were truncated to only include the coding regions of the GluN2 transmembrane domains.
conclusionsWe determined that viral packaging limit, transgene promoter and the presence of an intron within the transgene were all important factors that contributed to being able to successfully develop viral vectors designed to deliver and express GluN2 transgenes in a neuron specific manner. Because these vectors have been optimized to accommodate large open reading frames and in some cases contain an intron to facilitate expression of difficult to express transgenes, these viral vectors likely could be useful for delivering and expressing many large or difficult to express transgenes in a neuron specific manner.
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