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Stable STIM1 knockdown in human neural precursors derived from hESC line, H9

ABSTRACT: Stable STIM1 knockdown in human neural precursors derived from hESC line, H9

PROVIDER: PRJNA429650 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6464810.fastq.gz	Fastqsanger.gz
	SRR6464811.fastq.gz	Fastqsanger.gz
	SRR6464812.fastq.gz	Fastqsanger.gz
	SRR6464813.fastq.gz	Fastqsanger.gz
	SRR6464805.fastq.gz	Fastqsanger.gz

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Stable STIM1 knockdown in human neural precursors derived from hESC line, H9

Project description:Ca2 + signaling plays a significant role in development of the vertebrate nervous system where it regulates neurite growth as well as synapse and neurotransmitter specification (Rosenberg and Spitzer, 2011). Elucidating the role of Ca2 + signaling in neuronal development has been largely restricted to either small animal models or primary cultures. Here we derived human neural precursor cells (NPCs) from human embryonic stem cells to understand the functional significance of a less understood arm of calcium signaling, Store-operated Ca2+ entry or SOCE, in neuronal development. Human NPCs exhibited robust SOCE, which was significantly attenuated by expression of a stable shRNA-miR targeted towards the SOCE molecule STIM1. Along with the plasma membrane channel Orai, STIM is an essential component of SOCE in many cell types where it regulates gene expression. Therefore, we measured global gene expression in human NPCs with and without STIM1 knockdown. Interestingly, pathways down-regulated through STIM1 knockdown were related to cell proliferation and DNA replication processes whereas post-synaptic signaling was identified as an up-regulated process. To understand the functional significance of these gene expression changes we measured the self-renewal capacity of NPCs with STIM1 knockdown. These demonstrated significantly reduced neurosphere size and number as compared to control cells. Moreover, spontaneous differentiation towards the neuronal lineage was enhanced. These findings demonstrate that STIM1 mediated SOCE in human NPCs regulates gene expression changes, that in vivo are likely to physiologically modulate the self-renewal and differentiation of NPCs.

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ChIP-seq for Nanog in H9 hESC

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Gene expression analysis of H9 hESC derived cardiomyocytes (CM) and neuron stem cells (NSC)

Project description:In this study, we generated wildtype H9 hESC derived cardiomyocytes (CM) and neural stem cells (NSC) by in vitro differentiation. Global gene expression profiles were compared among undifferentiated H9 hESC and the derived CM and NSC. Comparison of global gene expression profiles of undifferentiated H9 hESC and the derived CM and NSC populations.

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Gene expression analysis of H9 hESC derived cardiomyocytes (CM) and neuron stem cells (NSC)

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Gene expression analysis of H9 hESC derived neuron stem cells (NSC) harboring pathogenic LRRK2 (G2019S) mutation

Project description:Genetic mutations on leucine-rich repeat kinase 2 (LRRK2) have been associated with an increased risk of Parkinson's disease. The Gly2019Ser (G2019S) mutation on LRRK2 gene is a relatively common cause of familial Parkinson's disease in Caucasian population. In this study, we generated H9 hESC harboring LRRK2 (G2019S) mutation by gene knockin. Wildtype and LRRK2 mutant hESC were differentiated into NSC using a chemically defined protocol.

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Global Gene Expression of H9 hESC with different passage numbers

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Single-cell RNA-seq of hESC (H9) lines exposed to a striatal differentiation protocol

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Analysis of the protein cargo of extracellular vesicles isolated from cultured line H9 human stem cells

Project description:Extracellular vesicles (EV) are secreted by nearly every mammalian cell type and contain a wealth of bioactive cargo capable of modulating target cell physiology and function though a variety of paracrine signaling mechanisms (Leavitt et al., 2019). Human embryonic stem cell (hESC)-derived extracellular vesicles (hESC-derived EV), were extracted from the hESC line H9 (WA09 Wicell Research Institute, Inc., Madison, WI). We analyzed the bioactive protein cargo to identify components that help to resolve radiation-induced injury to the lung in mice when injected in vivo.

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