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ABSTRACT: RNA sequencing of Colobanthus quitensis during during post-thaw recovery

PROVIDER: PRJNA1099400 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28638925_1.fastq.gz	Fastqsanger.gz
	SRR28638925_2.fastq.gz	Fastqsanger.gz
	SRR28638926_1.fastq.gz	Fastqsanger.gz
	SRR28638926_2.fastq.gz	Fastqsanger.gz
	SRR28638927_1.fastq.gz	Fastqsanger.gz

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Evaluation of transcriptomic changes in a Neuro2a cell line under extreme stress

Project description:Bulk poly(A) RNA-sequencing was performed to evaluate the transcriptomic profiles of Neuro2a cells exposed to extreme stress following thawing in liquid N2 storage. This RNA-seq experiment was conducted as a part of broader aim in our study to evaluate changes gene expression of stressed cells in the early stages of recovery at Passage 1 post-thaw (P1) and as cells progressed towards a healthy phenotype at Passage 6 post-thaw (P6). Briefly, RNA was extracted and purified from Neuro2a cells at P1 (n=4 for experimental; n=3 for controls) and at P6 (n=3/group); poly(A) RNA was isolated, fragmented, and converted into cDNA libraries. Libraries were sequenced on an Illumina NovaSeq 6000 instrument at an approximate depth of 40 million 150 bp paired-end reads per sample. Sequencing yielded an average of 38.99 million reads per sample with and average of 94.91% of reads having a quality score of at least 30 (Q30) and an average of 96.29% of reads mapping to the mouse reference genome.

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Insufficiency of Copper Ion Homeostasis Causes Freeze-Thaw Injury of Yeast Cells

Project description:Saccharomyces cerevisiae is exposed to freeze-thaw stress in commercial processes including frozen dough baking. The cell viability and fermentation activity after freeze-thaw were dramatically decreased due to freeze-thaw injury. Because freeze-thaw injury involves complex phenomena, the mechanisms of it are not fully understood. We attempted to analyze the mechanisms of freeze-thaw injury by indirect gene expression analysis during post-thaw incubation after freeze-thaw treatment using DNA microarray profiling. The results showed that a high frequency of the genes involved in the homeostasis of metal ions were up-regulated depending on the freezing period. The phenotype of the deletion mutants of the up-regulated genes extracted by indirect gene expression analysis was assessed. The deletion strains of the MAC1 and CTR1 genes involved in copper ion homeostasis exhibited freeze-thaw sensitivity, suggesting that copper ion homeostasis is required for freeze-thaw tolerance. Supplementation with copper ions during post-thaw incubation increased intracellular superoxide dismutase activity. Inverse correlated with intracellular superoxide dismutase activity, intracellular levels of reactive oxygen species were decreased. Moreover, cell viability increased by supplementation with copper ions under specific assessment conditions. This study suggested that insufficiency of copper ion homeostasis may be one of the causes of freeze-thaw injury.

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