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Purification of mitochondria from skeletal muscle tissue for transcriptomic analyses reveals localization of nuclear-encoded non-coding RNAs [Mitoplasts]

ABSTRACT: Purification of mitochondria from skeletal muscle tissue for transcriptomic analyses reveals localization of nuclear-encoded non-coding RNAs [Mitoplasts]

PROVIDER: PRJNA1156604 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30543030_1.fastq.gz	Fastqsanger.gz
	SRR30543030_2.fastq.gz	Fastqsanger.gz
	SRR30543031_1.fastq.gz	Fastqsanger.gz
	SRR30543031_2.fastq.gz	Fastqsanger.gz
	SRR30543032_1.fastq.gz	Fastqsanger.gz

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Purification of mitochondria from skeletal muscle tissue for transcriptomic analyses reveals localization of nuclear-encoded non-coding RNAs [SMvsMT]

Project description:Mitochondria are central to cellular function, particularly in metabolically active tissues such as skeletal muscle. Nuclear-encoded RNAs typically localise within the nucleus and cytosol but a small population may also translocate to subcellular compartments such as mitochondria. We aimed to investigate the nuclear-encoded RNAs that localise within the mitochondria of skeletal muscle tissue. Intact mitochondria were isolated via immunoprecipitation (IP) followed by enzymatic treatments (RNase-A and proteinase-K) optimised to remove transcripts located exterior to mitochondria, making it amenable for high-throughput transcriptomic sequencing. Whole-transcriptome RNA sequencing of enzymatically-purified mitochondria isolated by IP from skeletal muscle tissue showed a high degree of purity. In summary, we describe a novel, powerful sequencing approach applicable to animal and human tissues and cells that can facilitate the discovery of nuclear-encoded RNA transcripts localised within skeletal muscle mitochondria.

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