Sensing Self and Foreign Circular RNAs by Intron Identity
Open accessAbstract
Circular RNAs (circRNAs) are single-stranded RNAs that are joined head to tail with largely unknown functions. Here we show that transfection of purified in vitro generated circRNA into mammalian cells led to potent induction of innate immunity genes and confers protection against viral infection. The nucleic acid sensor RIG-I is necessary to sense foreign circRNA, and RIG-I and foreign circRNA co-aggregate in cytoplasmic foci. CircRNA activation of innate immunity is independent of a 5' triphosphate, double-stranded RNA structure, or the primary sequence of the foreign circRNA. Instead, self-nonself discrimination depends on the intron that programs the circRNA. Use of a human intron to express a foreign circRNA sequence abrogates immune activation, and mature human circRNA is associated with diverse RNA binding proteins reflecting its endogenous splicing and biogenesis. These results reveal innate immune sensing of circRNA and highlight introns-the predominant output of mammalian transcription-as arbiters of self-nonself identity.
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Transfection of cells with exogenous circRNA results in activation of antiviral gene products such as OAS, PKR, and RIG-I, and RIG-I is likely the principal component responsible for the cellular response against circRNA.
“It has recently been demonstrated that transfection of cells with exogenous circRNA results in the activation of antiviral gene products such as OAS, PKR, and RIG-I, and that RIG-I is likely to be the principal component responsible for the cellular response against circRNA28.”
From Engineering circular RNA for potent and stable translation in eukaryotic cells (2018) · cites this paper as [28]
The 3-fold induction of RIG-I mRNA observed was significantly lower than the 500-fold induction reported by Chen et al. in HeLa cells and was not sufficient to suppress protein translation and the stability of protein expression from circRNA in this cellular context.
“The 3-fold induction of RIG-I mRNA we observed was significantly lower than the 500-fold induction reported by Chen et al. in HeLa cells28 and was not sufficient to suppress protein translation and the stability of protein expression from circRNA in this cellular context (Fig. 6d).”
From Engineering circular RNA for potent and stable translation in eukaryotic cells (2018) · cites this paper as [28]
Unmodified circRNA has been shown to induce TLR/RIG-I-mediated innate immune response compared to unmodified linear mRNA.
“unmodified circRNA has been shown to induce TLR/RIG-I-mediated innate immune response compared to unmodified linear mRNA.225,226”
From Advances in COVID-19 mRNA vaccine development (2022) · cites this paper as [226]
Previous studies used circRNA purified by RNase R.
“Previous studies have used circRNA purified by RNase R (Chen et al. 2017).”
From RNA Circularization Diminishes Immunogenicity and Can Extend Translation Duration In Vivo (2019) · cites this paper as Chen et al. 2017
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- Authors
- Chen, Y. G., Kim, M. V., Chen, X., Batista, P. J., Aoyama, S., Wilusz, J. E., Iwasaki, A., & Chang, H. Y.
- Affiliations
- Stanford Medicine (US)
- Howard Hughes Medical Institute (US)
- University of Pennsylvania (US)
- Corresponding author
- Chang, Howard Y.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2017-07
- Volume
- 67
- Issue
- 2
- Pages
- 228-238.e5
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC5610545
- OpenAlex
- W2626686215
- Cited by
- 475
- References
- 39
- Citations by year
- 2026: 41 · 2025: 56 · 2024: 53 · 2023: 40 · 2022: 51 · 2021: 73 · 2020: 65 · 2019: 44 · 2018: 44 · 2017: 7
- FWCI
- 17.78
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Dean’s Fellowship and National Institutes of Health Postgraduate Training Program (2T32AR00742231)
- James Hudson Brown-Alexander Brown Coxe Fellowship
- Swedish Research Council International Postdoctoral Fellowship (VR-2016-06794)
- Damon Runyon Cancer Research Foundation (2090-11)
- National Institutes of Health Postgraduate Training Program (2T32AR00742231)
- Rita Allen Foundation Scholar
- Howard Hughes Medical Institute
- National Institutes of Health (R00-GM104166, R35-GM119735, R41AI120269, P50-HG007735, R01-HG004361)
- Parker Institute for Cancer Immunotherapy
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Circular RNAs in diseases
- Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- circular RNA, RIG-I, innate immunity, self/non-self discrimination, introns, viral infection protection, RNA-binding proteins, circRNA biogenesis, innate immune activation
- MeSH terms
- RAW 264.7 Cells, DEAD Box Protein 58, RNA, Circular, Animals, Base Sequence, Encephalitis Virus, Venezuelan Equine, Encephalomyelitis, Venezuelan Equine, HeLa Cells, Humans, Immune Tolerance, Immunity, Innate, Introns, Nucleic Acid Conformation, Protein Binding, Receptors, Immunologic, RNA, RNA Processing, Post-Transcriptional, RNA, Messenger, Transfection, RNA-Binding Proteins, Spliceosomes, Mice, Host-Pathogen Interactions, HEK293 Cells
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