RNAi-directed knockdown induces nascent transcript degradation and premature transcription termination in the nucleus

You, Jin Sam; Song, Zhenxing; Lin, Jiamei; Jia, Ruirui; Xia, Fei; Li, Zhengguo; Huang, Chuan

doi:10.1038/s41421-021-00297-8

Cited by 9 publications

(9 citation statements)

References 13 publications

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“…To reveal the subcellular distribution of circSfl and CdSfl, cellular fractionation was performed as previously described ( 43 , 47 ). Briefly, the circSfl stable cell line was treated with 500 μM CuSO 4 for 12 h to induce circSfl and CdSfl expression.…”

Section: Methodsmentioning

confidence: 99%

“…CLIP assays were performed as previously described ( 5 , 47 ). Briefly, cells were washed with ice-cold 1× PBS buffer twice, irradiated in a UV cross-linker (Lanyi, LYUV07-11) with 400 mJ/cm 2 at 254 nm on ice for 1 min, and incubated with RIPA buffer in the presence of RNase inhibitor (80 units/ml; Beyotime, R0102) and 1× Protease Inhibitor Cocktail (Beyotime, P1045) for 30 min on ice followed by 100 rounds of pipetting.…”

Section: Methodsmentioning

confidence: 99%

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eIF3j inhibits translation of a subset of circular RNAs in eukaryotic cells

Song

Lin

et al. 2022

Nucleic Acids Research

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Increasing studies have revealed that a subset of circular RNAs (circRNAs) harbor an open reading frame and can act as protein-coding templates to generate functional proteins that are closely associated with multiple physiological and disease-relevant processes, and thus proper regulation of synthesis of these circRNA-derived proteins is a fundamental cellular process required for homeostasis maintenance. However, how circRNA translation initiation is coordinated by different trans-acting factors remains poorly understood. In particular, the impact of different eukaryotic translation initiation factors (eIFs) on circRNA translation and the physiological relevance of this distinct regulation have not yet been characterized. In this study, we screened all 43 Drosophila eIFs and revealed the conflicting functions of eIF3 subunits in the translational control of the translatable circRNA circSfl: eIF3 is indispensable for circSfl translation, while the eIF3-associated factor eIF3j is the most potent inhibitor. Mechanistically, the binding of eIF3j to circSfl promotes the disassociation of eIF3. The C-terminus of eIF3j and an RNA regulon within the circSfl untranslated region (UTR) are essential for the inhibitory effect of eIF3j. Moreover, we revealed the physiological relevance of eIF3j-mediated circSfl translation repression in response to heat shock. Finally, additional translatable circRNAs were identified to be similarly regulated in an eIF3j-dependent manner. Altogether, our study provides a significant insight into the field of cap-independent translational regulation and undiscovered functions of eIF3.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

eIF3j inhibits translation of a subset of circular RNAs in eukaryotic cells

Song

Lin

et al. 2022

Nucleic Acids Research

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“…1 and 2 ). siRNA-mediated inhibition of nascent transcription by premature transcription termination was recently observed in Drosophila melanogaster ( 39 ). To analyze whether VINK siRNAs induce premature transcription termination or involve the repression of VINK promoter, we analyzed the recruitment of RNA polymerase II (Pol II) to the TSS, which would be unaffected by the induction of premature termination.…”

Section: Resultsmentioning

confidence: 98%

Small Interfering RNAs Targeting a Chromatin-Associated RNA Induce Its Transcriptional Silencing in Human Cells

et al. 2022

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“…CircRNAs play pathogenic or suppressive roles and might serve as therapeutic targets in various diseases. Strategies, such as RNAi, ASO, and CRISPR‐mediated transcript knockdown (Abudayyeh et al, 2017; Özcan et al, 2021; Pamudurti et al, 2020; Song, Jia, et al, 2021; You et al, 2021), can be designed to reduce circRNA expression in specific tissues, and there are a few of studies supporting these new therapeutic approaches: (i) intratumor injection of the siRNA against circNRIP1 remarkably suppressed gastric cancer growth in mouse models (X. Zhang, Wang, et al, 2019); (ii) In vivo tail vein injection of the ASO targeting circLONP2 dramatically reduced the penetrance of metastasis to foreign organs in colorectal carcinoma (K. Han, Wang, et al, 2020b); (iii) genetically engineered circBoule knockout mice and fruit flies, which exhibit reduced male fertility in response to stressed conditions, have been successfully constructed by depleting the flanking introns of the circularizing exons through the CRISPR‐Cas 9 system (Gao et al, 2020).…”

Section: Therapeutic Strategies For Circular Rna Silencing and Overex...mentioning

confidence: 99%

Circular RNA in disease: Basic properties and biomedical relevance

et al. 2022

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Circular RNAs (circRNAs) represent a class of covalently closed RNA molecules with great diversity in molecular features, functions, and regulatory mechanisms. Emerging advances in our understanding of circRNA biogenesis, nuclear export, and stability control have been made very recently. In particular, novel roles of circRNAs in diverse human diseases are increasingly recognized. Various circRNAs have been found to affect many disease-relevant pathways through a diverse array of mechanisms, including forming R-loops, sponging miRNAs or proteins, and translating functional proteins, resulting in different pathological phenotypes. This recent progress calls for a revised view of circRNAs in diseases threatening the lives and health of humans. In this review, we focus on the recently described functional relevance of diseaseassociated circRNAs as well as the potential of circRNAs in diverse clinical applications. This article is categorized under: RNA in Disease and Development > RNA in Disease Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs K E Y W O R D S circular RNA, clinical application, nonimmunological disease 1 | INTRODUCTION Circular RNAs (circRNAs) are noncanonical splicing products with covalently closed loop structures. Comprehensive studies of circRNAs have transformed the view of circRNAs from accidental by-products of mis-splicing to stable and evolutionarily conserved molecules, which are specifically expressed in various cells, tissues, or developmental stages of eukaryotes, prokaryotes, and viruses (

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RNAi-directed knockdown induces nascent transcript degradation and premature transcription termination in the nucleus

Cited by 9 publications

References 13 publications

eIF3j inhibits translation of a subset of circular RNAs in eukaryotic cells

eIF3j inhibits translation of a subset of circular RNAs in eukaryotic cells

Small Interfering RNAs Targeting a Chromatin-Associated RNA Induce Its Transcriptional Silencing in Human Cells

Circular RNA in disease: Basic properties and biomedical relevance

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