Enhancer-Driven Gene Expression (EDGE) Enables the Generation of Viral Vectors Specific to Neuronal Subtypes

Nair, Rajeevkumar Raveendran; Blankvoort, Stefan; Lagartos, Maria Jose; Kentros, Clifford

doi:10.1016/j.isci.2020.100888

Cited by 46 publications

(32 citation statements)

References 82 publications

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“…Subsequently, the mCherry sequence was synthesized and cloned in an inverted orientation into EcoR1 and BamH1 sites in pAAV-CMV-FLEX-MCS-WPRE to make pAAV CMV-FLEX-mCherry-WPRE. AAV-CMV-FLEX-mCherry was recovered from pAAV CMV-FLEX-mCherry-WPRE as described elsewhere ( Nair et al, 2020 ; Nilssen et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

Local projections of layer Vb-to-Va are more prominent in lateral than in medial entorhinal cortex

Ohara

Blankvoort

Nair

et al. 2021

eLife

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The entorhinal cortex, in particular neurons in layer V, allegedly mediate transfer of information from the hippocampus to the neocortex, underlying long-term memory. Recently, this circuit has been shown to comprise a hippocampal output recipient layer Vb and a cortical projecting layer Va. With the use of in vitro electrophysiology in transgenic mice specific for layer Vb, we assessed the presence of the thus necessary connection from layer Vb-to-Va in the functionally distinct medial (MEC) and lateral (LEC) subdivisions; MEC, particularly its dorsal part, processes allocentric spatial information, whereas the corresponding part of LEC processes information representing elements of episodes. Using identical experimental approaches, we show that connections from layer Vb-to-Va neurons are stronger in dorsal LEC compared with dorsal MEC, suggesting different operating principles in these two regions. Although further in vivo experiments are needed, our findings imply a potential difference in how LEC and MEC mediate episodic systems-consolidation.

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Section: Methodsmentioning

confidence: 99%

Local projections of layer Vb-to-Va are more prominent in lateral than in medial entorhinal cortex

Ohara

Blankvoort

Nair

et al. 2021

eLife

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“…Recent advances in viral vectors that enable targeting specific neurons are likely essential for subsequent studies. These include viral tools targeting neurons with specific projections ( Inoue et al, 2015 ; Oyama et al, 2021 ), or specific interneurons in non-human primates ( Dimidschstein et al, 2016 ; Vormstein-Schneider et al, 2020 ; Mich et al, 2021 ), as well as enhancer-based viral vectors targeting entorhinal excitatory cells ( Nair et al, 2020 ). The cell-type-based global map provided in this study will accelerate the application of these genetic tools in monkeys for better understanding of the role of EC in memory and navigation and will further the translational potential of the experimental observations in monkeys.…”

Section: Discussionmentioning

confidence: 99%

Laminar Organization of the Entorhinal Cortex in Macaque Monkeys Based on Cell-Type-Specific Markers and Connectivity

Ohara

Yoshino

Kimura

et al. 2021

Front. Neural Circuits

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The entorhinal cortex (EC) is a major gateway between the hippocampus and telencephalic structures, and plays a critical role in memory and navigation. Through the use of various molecular markers and genetic tools, neuron types constituting EC are well studied in rodents, and their layer-dependent distributions, connections, and functions have also been characterized. In primates, however, such cell-type-specific understandings are lagging. To bridge the gap between rodents and primates, here we provide the first cell-type-based global map of EC in macaque monkeys. The laminar organization of the monkey EC was systematically examined and compared with that of the rodent EC by using immunohistochemistry for molecular markers which have been well characterized in the rodent EC: reelin, calbindin, and Purkinje cell protein 4 (PCP4). We further employed retrograde neuron labeling from the nucleus accumbens and amygdala to identify the EC output layer. This cell-type-based approach enabled us to apply the latest laminar definition of rodent EC to monkeys. Based on the similarity of the laminar organization, the monkey EC can be divided into two subdivisions: rostral and caudal EC. These subdivisions likely correspond to the lateral and medial EC in rodents, respectively. In addition, we found an overall absence of a clear laminar arrangement of layer V neurons in the rostral EC, unlike rodents. The cell-type-based architectural map provided in this study will accelerate the application of genetic tools in monkeys for better understanding of the role of EC in memory and navigation.

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“…However, this significantly reduces the packaging capacity of AAV vectors. The advanced strategy of substituting large promoters with unique enhancer sequences to drive transgene expression selectively in neurons in a restricted brain area was tested recently and showed great potential in combination with AAV vector as well [ 155 , 156 , 157 ]. Similar approaches employed for description and characterization of specific astrocytic enhancers have great potential for developing methods for highly selective regulation of astrocytes in specific regions or across the brain.…”

Section: Discussionmentioning

confidence: 99%

Genetic Constructs for the Control of Astrocytes’ Activity

Borodinova

Balaban

Bezprozvanny

et al. 2021

Cells

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In the current review, we aim to discuss the principles and the perspectives of using the genetic constructs based on AAV vectors to regulate astrocytes’ activity. Practical applications of optogenetic approaches utilizing different genetically encoded opsins to control astroglia activity were evaluated. The diversity of astrocytic cell-types complicates the rational design of an ideal viral vector for particular experimental goals. Therefore, efficient and sufficient targeting of astrocytes is a multiparametric process that requires a combination of specific AAV serotypes naturally predisposed to transduce astroglia with astrocyte-specific promoters in the AAV cassette. Inadequate combinations may result in off-target neuronal transduction to different degrees. Potentially, these constraints may be bypassed with the latest strategies of generating novel synthetic AAV serotypes with specified properties by rational engineering of AAV capsids or using directed evolution approach by searching within a more specific promoter or its replacement with the unique enhancer sequences characterized using modern molecular techniques (ChIP-seq, scATAC-seq, snATAC-seq) to drive the selective transgene expression in the target population of cells or desired brain regions. Realizing these strategies to restrict expression and to efficiently target astrocytic populations in specific brain regions or across the brain has great potential to enable future studies.

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Enhancer-Driven Gene Expression (EDGE) Enables the Generation of Viral Vectors Specific to Neuronal Subtypes

Cited by 46 publications

References 82 publications

Local projections of layer Vb-to-Va are more prominent in lateral than in medial entorhinal cortex

Local projections of layer Vb-to-Va are more prominent in lateral than in medial entorhinal cortex

Laminar Organization of the Entorhinal Cortex in Macaque Monkeys Based on Cell-Type-Specific Markers and Connectivity

Genetic Constructs for the Control of Astrocytes’ Activity

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