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Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [Ago2-CLEAR-CLIP]

ABSTRACT: Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [Ago2-CLEAR-CLIP]

PROVIDER: PRJNA1087698 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28342437.fastq.gz	Fastqsanger.gz
	SRR28342438.fastq.gz	Fastqsanger.gz
	SRR28342439.fastq.gz	Fastqsanger.gz
	SRR28342440.fastq.gz	Fastqsanger.gz

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Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [Ago2-CLEAR-CLIP]

Project description:RNA interference (RNAi) is a fundamental strategy for gene silencing via small interfering RNAs (siRNAs) that program Argonaute (Ago) effectors, and is related to endogenous silencing via microRNAs (miRNAs). Although most efforts focus on factors that are required for miRNA/RNAi function, biological systems are typically subject to repression. Using molecular genetic approaches, we uncover ALAS1, the first enzyme in the heme biosynthesis pathway, as a repressor of miRNA accumulation in cells and in mice. This was non-intuitive, since heme is known to play a positive role as a cofactor for the nuclear miRNA processing machinery. Instead, we show that ALAS1 (but not other core heme biosynthesis factors) limits RISC assembly and activity. Notably, ALAS1 is the target of one the few current FDA-approved siRNA drugs (Givosiran). Our data have implications for side effects of Givosiran, but also for RNAi therapies in general. We provide evidence that depletion of ALAS1 enhances siRNA-mediated knockdown, suggesting that Givosiran is a viable RNAi adjuvant.

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Project description:We deciphered Ago2:RNA interactions in post-mortem human heart tissues using crosslinking immunoprecipitation coupled with high-throughput sequencing (HITS-CLIP) to generate the first transcriptome-wide map of miR targeting events in human myocardium, detecting 4000 cardiac Ago2 binding sites across >2200 target transcripts.

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miRBShunter: a pipeline to identify miRNA binding sites from Ago2-CLIP data by de novo motif finding

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Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [HEK293T_sRNA-seq]

Project description:Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [HEK293T_sRNA-seq]

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Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [hepatocyte_sRNA-seq]

Project description:Noncanonical role of ALAS1 as a heme-independent inhibitor of small RNA–mediated silencing [hepatocyte_sRNA-seq]

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