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The effect of 22-bulge of shRNAs on DICER cleavage in human cells

ABSTRACT: The effect of 22-bulge of shRNAs on DICER cleavage in human cells

PROVIDER: PRJNA757441 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15600182_1.fastq.gz	Fastqsanger.gz
	SRR15600182_2.fastq.gz	Fastqsanger.gz
	SRR15600183_1.fastq.gz	Fastqsanger.gz
	SRR15600183_2.fastq.gz	Fastqsanger.gz
	SRR15600184_1.fastq.gz	Fastqsanger.gz

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The effect of 22-bulge of shRNAs on DICER cleavage in human cells

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The effect of 22-bulge of pre-miRNAs on DICER cleavage in human cells

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The loop position of shRNAs and pre-miRNAs is critical for the accuracy of Dicer processing in vivo

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Two-motif model illuminates DICER's cleavage preferences [DICER-mutants]

Project description:In humans, DICER is a key regulator of gene expression in animals through its production of miRNAs and siRNAs by processing miRNA precursors (pre-miRNAs), short-hairpin RNAs (shRNAs), and long double-stranded RNAs (dsRNAs). To advance our understanding of this process, we employed high-throughput assays using various shRNA variants and both wild-type and mutant DICER (DICERΔdsRBD). Our analysis revealed that DICER predominantly cleaves shRNAs at two positions, specifically at 21 (DC21) and 22 (DC22) nucleotides from the 5'-end. Our investigation identified two independent motifs, mWCU and YCR, that determine whether DICER cleaves at DC21 or DC22, depending on their locations in shRNA/pre-miRNAs. These motifs can work together or independently to determine the cleavage sites of DICER. Furthermore, our findings indicate that dsRBD enhances cleavage at DC21, and mWCU strengthens the interaction between dsRBD and RNA, leading to an even greater enhancement of DC21 cleavage. Conversely, YCR functions independently of dsRBD. Our study proposes a two-motif model that sheds light on the intricate regulatory mechanisms involved in gene expression by elucidating how DICER recognizes its substrate, providing valuable insights into this critical biological process.

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Project description:The high-throughput DICER cleavage assays were conducted shRNA variants containing different sequences. We showcase a comprehensive cleavage activity of DICER on different shRNAs containing different secondary structures.

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