Multiplexed CRISPRi Reveals a Transcriptional Switch Between KLF Activators and Repressors in the Maturing Neocortex

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 8, 2025

Abstract A critical phase of mammalian brain development takes place after birth. Neurons the mouse neocortex undergo dramatic changes in their morphology, physiology, and synaptic connections during first postnatal month, while properties immature neurons, such as capacity for robust axon outgrowth, are lost. The genetic epigenetic programs controlling prenatal well studied, but our understanding transcriptional mechanisms that regulate neuronal maturation is comparatively lacking. By integrating chromatin accessibility gene expression data from two subtypes neocortical pyramidal neurons neonatal maturing brain, we predicted a role Krüppel-Like Factor (KLF) family Transcription Factors developmental regulation neonatally expressed genes. Using multiplexed CRISPR Interference (CRISPRi) knockdown strategy, found shift KLF activators (Klf6, Klf7) to repressors (Klf9, Klf13) early functions ‘switch’ activate, then repress set shared targets with cytoskeletal including Tubb2b Dpysl3 . We demonstrate this switch buffered by redundancy between paralogs, which CRISPRi strategy equipped overcome study. Our results indicate competition within regulates conserved component program may underlie loss intrinsic growth neurons. This could facilitate transition refinement required stabilize mature circuits.

Language: Английский

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Multiplexed CRISPRi Reveals a Transcriptional Switch Between KLF Activators and Repressors in the Maturing Neocortex

Published: April 30, 2025

A critical phase of mammalian brain development takes place after birth. Neurons the mouse neocortex undergo dramatic changes in their morphology, physiology, and synaptic connections during first postnatal month, while properties immature neurons, such as capacity for robust axon outgrowth, are lost. The genetic epigenetic programs controlling prenatal well studied, but our understanding transcriptional mechanisms that regulate neuronal maturation is comparatively lacking. By integrating chromatin accessibility gene expression data from two subtypes neocortical pyramidal neurons neonatal maturing brain, we predicted a role Krüppel-Like Factor (KLF) family Transcription Factors developmental regulation neonatally expressed genes. Using multiplexed CRISPR Interference (CRISPRi) knockdown strategy, found shift KLF activators (Klf6, Klf7) to repressors (Klf9, Klf13) early functions ‘switch’ activate, then repress set shared targets with cytoskeletal including Tubb2b Dpysl3 . We demonstrate this switch buffered by redundancy between paralogs, which CRISPRi strategy equipped overcome study. Our results indicate competition within regulates conserved component program may underlie loss intrinsic growth neurons. This could facilitate transition refinement required stabilize mature circuits.

Language: Английский

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Multiplexed CRISPRi Reveals a Transcriptional Switch Between KLF Activators and Repressors in the Maturing Neocortex

Published: April 30, 2025

A critical phase of mammalian brain development takes place after birth. Neurons the mouse neocortex undergo dramatic changes in their morphology, physiology, and synaptic connections during first postnatal month, while properties immature neurons, such as capacity for robust axon outgrowth, are lost. The genetic epigenetic programs controlling prenatal well studied, but our understanding transcriptional mechanisms that regulate neuronal maturation is comparatively lacking. By integrating chromatin accessibility gene expression data from two subtypes neocortical pyramidal neurons neonatal maturing brain, we predicted a role Krüppel-Like Factor (KLF) family Transcription Factors developmental regulation neonatally expressed genes. Using multiplexed CRISPR Interference (CRISPRi) knockdown strategy, found shift KLF activators (Klf6, Klf7) to repressors (Klf9, Klf13) early functions ‘switch’ activate, then repress set shared targets with cytoskeletal including Tubb2b Dpysl3 . We demonstrate this switch buffered by redundancy between paralogs, which CRISPRi strategy equipped overcome study. Our results indicate competition within regulates conserved component program may underlie loss intrinsic growth neurons. This could facilitate transition refinement required stabilize mature circuits.

Language: Английский

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Asynchronous development of the mouse auditory cortex is driven by hemispheric identity and sex

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: April 17, 2025

Lateralized auditory processing is essential for specialized functions such as speech processing, typically dominated by the Left Auditory Cortex (ACx) in humans. Hemispheric specializations also occur adult mouse ACx, but their developmental origins are unclear. Our study finds that and Right ACx mice reach milestones at different ages. Thalamocortical responses maturation of synaptic dynamics develop earlier than Left. We show this timing offset predicts hemisphere-dependent differences sensory-driven plasticity. Juvenile tone exposure specific times results imbalanced frequency representations ACx. Additionally, sex influences plasticity; female plasticity occurs before male aligns with findings demonstrate hemispheric identity drive asynchronous development contribute to functional sensory cortices.

Language: Английский

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Cell-extrinsic controls over neocortical neuron fate and diversity

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 16, 2024

Cellular diversity in the neocortex emerges gradually during prenatal and postnatal development. While environmental interactions occur this extended maturation period, impact of extrinsic cues on determining fate distinct neuron types remains unknown. To address question, we exposed developing neocortical cells to various conditions examined how affects cell diversity. Our developmental analyses reveal a hierarchical molecular program which class-distinguishing features emerge first, followed by subclass- type-related characteristics, with paces among populations. Environmental contribution was assessed vivo, using genetically modified mice models position or innervation are altered, vitro two-dimensional cultures. Acquisition cellular identity remained stable across vivo models. In contrast, glutamatergic neurons showed decreased expression identity-defining genes, reduced alterations canonical cortical connectivity. were restored towards values organotypic slice These findings population-specific responses highlight role extracellular context shaping maturing neocortex.

Language: Английский

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