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Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs

ABSTRACT: Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs

PROVIDER: PRJNA692618 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13448602_1.fastq.gz	Fastqsanger.gz
	SRR13448602_2.fastq.gz	Fastqsanger.gz
	SRR13448603_1.fastq.gz	Fastqsanger.gz
	SRR13448603_2.fastq.gz	Fastqsanger.gz
	SRR13448604_1.fastq.gz	Fastqsanger.gz

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Robust RNA editing via recruitment of endogenous ADARs using circular guide RNAs

Project description:We engineered circular ADAR recruiting guide RNAs (cadRNAs) that efficiently recruit endogenous ADARs to edit specific sites on target RNA

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ADARs act as potent regulators of circular transcriptome in cancer

Project description:Circular RNAs (circRNAs) are produced by head-to-tail back-splicing which is mainly facilitated by base-pairing of reverse complementary matches (RCMs) in circRNA flanking introns. Adenosine deaminases acting on RNA (ADARs) are known to bind double-stranded RNAs for adenosine to inosine (A-to-I) RNA editing. Here we characterize ADARs as potent regulators of circular transcriptome by identifying over a thousand of circRNAs regulated by ADARs in a bidirectional manner through and beyond their editing function. We found that editing can stabilize or destabilize secondary structures formed between RCMs via correcting A:C mismatches to I(G)-C pairs or creating I(G).U wobble pairs, respectively. We provided experimental evidence that editing also favors the binding of RNA-binding proteins such as PTBP1 to regulate back-splicing. These ADARs-regulated circRNAs which are ubiquitously expressed in multiple types of cancers, demonstrate high functional relevance to cancer. Our findings support a hitherto unappreciated bidirectional regulation of circular transcriptome by ADARs and highlights the complexity of cross-talk in RNA processing and its contributions to tumorigenesis.

2022-01-25 | GSE164681 | GEO

Efficient and precise editing of endogenous transcripts with SNAP-tagged ADARs

Project description:Molecular tools to target RNA site‐specifically allow recoding of RNA information and processing. SNAP‐tagged deaminases, guided by a chemically stabilized guideRNA, enable the simultaneous editing of targeted adenosine to inosine in several endogenous transcripts, with high efficiency (up to 90%), high potency, sufficient duration, and high precision. We applied SNAP‐ADARs for the efficient and concurrent editing of two disease‐relevant signaling transcripts, KRAS and STAT1. We also show improved performance compared to the recently described Cas13b-ADAR.

2018-06-01 | GSE112787 | GEO

ADARs regulate small RNAs mapped to edited sequences

Project description:Cellular RNAs containing double-stranded RNA (dsRNA) structures are subject to A-to-I RNA editing by the adenosine deaminases that act on RNA (ADARs). While A-to-I editing can alter mRNA coding potential, most editing is observed in non-coding sequences, the function of which remains poorly characterized. To correlate small RNA population with expression patterns of ADARs and hyperedited RNAs (editing-enriched regions: EERs) defined and characterized in a separate RNAseq analysis, we re-analyzed existing smallRNAseq datasets of a wildtype strain and a strain lacking ADARs (adr-1;adr-2). Analysis of primary siRNAs from mixed-stage worms revealed that ADARs impact siRNA biogenesis from EERs. We then analyzed primary and secondary RNAs mapping to EERs from embryo-stage or L4-stage worms and observed that ADAR effects on siRNA levels are dependent on developmental stage.

2018-02-23 | GSE89765 | GEO

Recruiting Endogenous ADARs with Antisense Oligonucleotides to Reprogram the Transcriptome

Project description:Huge efforts are made to engineer safe and efficient genome editing tools. An alternative might be the harnessing of ADAR-mediated RNA editing. We now present the engineering of chemically optimized antisense oligonucleotides that recruit endogenous human ADARs to edit endogenous transcripts in a simple and programmable way, an approach we refer to as RESTORE. Notably, RESTORE was markedly precise, and there was no evidence for perturbation of the natural editing homeostasis. We applied RESTORE to a panel of standard human cell lines, but also to several human primary cells including hepatocytes. In contrast to other RNA and DNA editing strategies, this approach requires only the administration of an oligonucleotide, circumvents the ectopic expression of proteins, and thus represents an attractive platform for drug development. In this respect we have shown the repair of the PiZZ mutation causing α1-antitrypsin deficiency and the editing of phosphotyrosine 701 in STAT1.

2018-12-01 | GSE121573 | GEO

ADARs act as potent regulators of circular transcriptome in cancer

Project description:ADARs act as potent regulators of circular transcriptome in cancer

| PRJNA691548 | ENA

Recruiting Endogenous ADARs with Antisense Oligonucleotides to Reprogram the Transcriptome

Project description:Recruiting Endogenous ADARs with Antisense Oligonucleotides to Reprogram the Transcriptome

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Effects of ADARs on small RNA processing pathways in C. elegans

Project description:Adenosine deaminases that act on RNA (ADARs) are RNA editing enzymes that convert adenosine to inosine in double-stranded RNA (dsRNA). To evaluate effects of ADARs on small RNAs that derive from dsRNA precursors, we performed deep-sequencing, comparing small RNAs from wildtype and ADAR mutant C. elegans. While editing in small RNAs was rare, at least 40% of microRNAs had altered levels in at least one ADAR mutant strain, and miRNAs with significantly altered levels had mRNA targets with correspondingly affected levels. About 40% of siRNAs derived from endogenous genes (endo-siRNAs) also had altered levels in at least one mutant strain, including 63% of Dicer-dependent endo-siRNAs. The 26G class of endo-siRNAs was significantly affected by ADARs, and many altered 26G loci had intronic reads, and histone modifications associated with transcriptional silencing. Our data indicate ADARs, through both direct and indirect mechanisms, are important for maintaining wildtype levels of many small RNAs in C. elegans.

2012-04-05 | GSE28888 | GEO

ADARs affect the biogenesis of many small RNAs in C. elegans, but few mature small RNAs are edited

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Efficient and precise editing of endogenous transcripts with SNAP-tagged ADARs

Project description:Efficient and precise editing of endogenous transcripts with SNAP-tagged ADARs

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