Pluto Bioinformatics

GSE104442: Alteration of transcriptome by recombinant ncRNA agents

Bulk RNA sequencing

Purpose: Identify whether recombinant ncRNAs are recognized by mammalian cells and processed into mature microRNAs, thus leading to the alteration of downstream target genes. In addition, we also explored whether the alteration of known or novel target genes are Dicer dependent.; Methods: HEK293T (Dicer-WT) and 424 (Dicer-KO) cell lines were treated with 20nM of control tRNA (MSA), nCAR/miR-34a-5p, or nCAR/miR-34a-3p in triplicate.; Results: Read counts of known mRNAs were analyzed using a RSEM-EdgeR workflow. Sequence reads were mapped to the human reference genome (hg19) by BWA, read counts for individual genes were computed using RSEM, and EdgeR was used to analyze significant (log2FC > 1.2 or log2FC < 0.8. P-value < 0.05 and log2CPM > 5) differentially expressed mRNAs.; Conclusions: Upon transfection of recombinant RNA agents in mammalian cells, known and novel target genes were regulated in a Dicer dependent manner (nCAR/miR-34a-5p) and also in a Dicer independent manner (nCAR/miR-124a-3p). SOURCE: Pui Yan Ho ( - UC Davis

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