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Engineered mischarged transfer RNAs for correcting pathogenic missense mutations [MSR-seq]

ABSTRACT: Engineered mischarged transfer RNAs for correcting pathogenic missense mutations [MSR-seq]

PROVIDER: PRJNA991918 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25181423_1.fastq.gz	Fastqsanger.gz
	SRR25181423_2.fastq.gz	Fastqsanger.gz
	SRR25181424_1.fastq.gz	Fastqsanger.gz
	SRR25181424_2.fastq.gz	Fastqsanger.gz
	SRR25181425_1.fastq.gz	Fastqsanger.gz

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Engineered mischarged transfer RNAs for correcting pathogenic missense mutations [MSR-seq]

Project description:Missense mutations account for nearly 50% of pathogenic mutations in human genetic diseases, most lack effective treatments. Gene therapies, CRISPR-based gene editing, and RNA therapies including transfer RNA (tRNA) modalities are common strategies for potential treatments of genetic diseases. However, reported tRNA therapies are for nonsense mutations, how tRNAs can be engineered to correct missense mutations have not been explored. Here, we describe missense correcting tRNAs (mc-tRNAs) as a potential therapeutic modality for correcting pathogenic missense mutations. Mc-tRNAs are engineered tRNAs that are charged with one amino acid and read codons of another amino acid in translation in human cells. We first developed a series of fluorescence protein (FP)-based reporters that indicate successful correction of missense mutations via restoration of fluorescence signals. We engineered mc-tRNAs that effectively corrected Serine and Arginine missense mutations in the reporters and confirmed the amino acid substitution by protein mass spectrometry and mc-tRNA expression by tRNA sequencing. We examined the transcriptome response to the expression of mc-tRNAs and found some mc-tRNAs induced minimum transcriptomic changes. Furthermore, we applied an Arg-tRNAGln(CUG) mc-tRNA to rescue the autolytic activity of a pathogenic CAPN3 Arg-to-Gln mutant involved in limb-girdle muscular dystrophy type 2A. These results establish a versatile pipeline for mc-tRNA engineering and demonstrate the potential of mc-tRNA as an alternative therapeutic platform for the treatment of genetic disorders.

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Engineered mischarged transfer RNAs for correcting pathogenic missense mutations

Project description:Engineered mischarged transfer RNAs for correcting pathogenic missense mutations

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Engineered mischarged transfer RNAs for correcting pathogenic missense mutations [mRNA-seq]

Project description:Engineered mischarged transfer RNAs for correcting pathogenic missense mutations [mRNA-seq]

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