Engineering circular RNA expression systems to minimize contaminating linear RNA byproducts.
Fields CJ, Fujiwara R, Wright BW, Stringer BW, Conn SJ, Wilusz JE
bioRxiv [Preprint]. 2026 Sep 11:2026.09.07.749908. doi: 10.64898/2026.09.07.749908.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 262612 | pcDNA3.1(+) circRNA Mini Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262613 | pcDNA3.1(+) ZKSCAN1 Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262614 | pcDNA3.1(+) Laccase2 Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262615 | pcDNA3.1(+) POLR2A Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262616 | pcDNA3.1(+) ciRS-7 Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262617 | pcDNA3.1(+) IGF2BP1 Scarless MCS | Expression plasmid for expressing scarless circular RNAs of a desired sequence in mammalian cells |
| 262618 | pcDNA3.1(+) Tornado PaqCI MCS | Expression plasmid for expressing circular RNAs of a desired sequence in mammalian cells |
| 262619 | pcDNA3.1(+) circRNA Mini-Split eGFP | Expresses a scarred eGFP circular RNA containing the EMCV IRES in mammalian cells using mini ZKSCAN1 introns |
| 262620 | pcDNA3.1(+) circRNA Mini Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using mini ZKSCAN1 introns |
| 262621 | pcDNA3.1(+) ZKSCAN1 Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ZKSCAN1 introns |
| 262622 | pcDNA3.1(+) Laccase2 Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using Laccase2 introns |
| 262623 | pcDNA3.1(+) POLR2A Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using POLR2A introns |
| 262624 | pcDNA3.1(+) ciRS-7 Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ciRS-7 (CDR1as) introns |
| 262625 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns |
| 262626 | pcDNA3.1(+) Tornado PaqCI-Split eGFP | Expresses a scarred eGFP circular RNA containing the EMCV IRES in mammalian cells using the Tornado platform |
| 262627 | pcDNA3.1(+) circRNA Mini-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262628 | pcDNA3.1(+) circRNA Mini Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262629 | pcDNA3.1(+) ZKSCAN1 Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262630 | pcDNA3.1(+) Laccase2 Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262631 | pcDNA3.1(+) POLR2A Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262632 | pcDNA3.1(+) ciRS-7 Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262633 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP Delta Repeat | Control eGFP circular RNA expression plasmid that fails to backsplice due to deletion of downstream intronic repeat |
| 262634 | pcDNA3.1(+) ciRS-7 Scarless-Split eGFP (Laccase2 ss) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ciRS-7 (CDR1as) introns with Laccase2 splice sites |
| 262635 | pcDNA3.1(+) ciRS-7 Scarless-Split eGFP (ZKSCAN1 ss) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ciRS-7 (CDR1as) introns with ZKSCAN1 splice sites |
| 262636 | pcDNA3.1(+) circRNA Mini-ciRS-7 | Expresses scarred ciRS-7 (CDR1as) circRNA in mammailian cells using mini ZKSCAN1 introns |
| 262637 | pcDNA3.1(+) circRNA Mini Scarless-ciRS-7 | Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using mini ZKSCAN1 introns |
| 262638 | pcDNA3.1(+) ZKSCAN1 Scarless-ciRS-7 | Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using ZKSCAN1 introns |
| 262639 | pcDNA3.1(+) Laccase2 Scarless-ciRS-7 | Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using Laccase2 introns |
| 262640 | pcDNA3.1(+) POLR2A Scarless-ciRS-7 | Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using POLR2A introns |
| 262641 | pcDNA3.1(+) Tornado PaqCI-ciRS-7 | Expresses scarred ciRS-7 (CDR1as) circRNA in mammailian cells using the Tornado platform |
| 262642 | pcDNA3.1(+) ciRS-7 Scarless-ciRS-7 | Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using ciRS-7 (CDR1as) introns |
| 262643 | pcDNA3.1(+) circRNA Mini-ZKSCAN1 Exons 2-3 | Expresses scarred circZKSCAN1(2,3) circRNA in mammailian cells using mini ZKSCAN1 introns |
| 262644 | pcDNA3.1(+) circRNA Mini Scarless-ZKSCAN1 Exons 2-3 | Expresses scarless circZKSCAN1(2,3) circRNA in mammailian cells using mini ZKSCAN1 introns |
| 262645 | pcDNA3.1(+) ZKSCAN1 Scarless-ZKSCAN1 Exons 2-3 | Expresses scarless circZKSCAN1(2,3) circRNA in mammailian cells using ZKSCAN1 introns |
| 262646 | pcDNA3.1(+) Laccase2 Scarless-ZKSCAN1 Exons 2-3 | Expresses scarless circZKSCAN1(2,3) circRNA in mammailian cells using Laccase2 introns |
| 262647 | pcDNA3.1(+) POLR2A Scarless-ZKSCAN1 Exons 2-3 | Expresses scarless circZKSCAN1(2,3) circRNA in mammailian cells using POLR2A introns |
| 262648 | pcDNA3.1(+) ciRS-7 Scarless-ZKSCAN1 Exons 2-3 | Expresses scarless circZKSCAN1(2,3) circRNA in mammailian cells using ciRS-7 (CDR1as) introns |
| 262649 | pcDNA3.1(+) Tornado PaqCI-ZKSCAN1 Exons 2-3 | Expresses scarred ZKSCAN1(2,3) circRNA in mammailian cells using the Tornado platform |
| 262650 | pcDNA3.1(+) circRNA Mini-NFASC Exons 26-27 | Expresses scarred circNFASC(26,27) circRNA in mammailian cells using mini ZKSCAN1 introns |
| 262651 | pcDNA3.1(+) circRNA Mini Scarless-NFASC Exons 26-27 | Expresses scarless circNFASC(26,27) circRNA in mammailian cells using mini ZKSCAN1 introns |
| 262652 | pcDNA3.1(+) ZKSCAN1 Scarless-NFASC Exons 26-27 | Expresses scarless circNFASC(26,27) circRNA in mammailian cells using ZKSCAN1 introns |
| 262653 | pcDNA3.1(+) Laccase2 Scarless-NFASC Exons 26-27 | Expresses scarless circNFASC(26,27) circRNA in mammailian cells using Laccase2 introns |
| 262654 | pcDNA3.1(+) POLR2A Scarless-NFASC Exons 26-27 | Expresses scarless circNFASC(26,27) circRNA in mammailian cells using POLR2A introns |
| 262655 | pcDNA3.1(+) ciRS-7 Scarless-NFASC Exons 26-27 | Expresses scarless circNFASC(26,27) circRNA in mammailian cells using ciRS-7 (CDR1as) introns |
| 262656 | pcDNA3.1(+) Tornado PaqCI-NFASC Exons 26-27 | Expresses scarred NFASC(26,27) circRNA in mammailian cells using the Tornado platform |
| 262657 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Upstream miR-92a) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a miR-92a site in upstream intron |
| 262658 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Upstream miR-92a Mut) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a mutant miR-92a site in upstream intron |
| 262659 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Downstream miR-92a) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a miR-92a site in downstream intron |
| 262660 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Downstream miR-92a Mut) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a mutant miR-92a site in downstream intron |
| 262661 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Up & Downstream miR-92a) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a miR-92a site in up- and downstream introns |
| 262662 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (Up & Downstream miR-92a Mut) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns that have a mutant miR-92a site in up- anddownstream introns |
| 262663 | pcDNA3.1(+) ZKSCAN1 Scarless-Split eGFP (mascRNA WT) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ZKSCAN1 introns. Instead of a poly(A) signal, the primary transcript ends in WT mascRNA to destabilize linear RNAs. |
| 262664 | pcDNA3.1(+) ZKSCAN1 Scarless-Split eGFP (mascRNA Mut 7) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ZKSCAN1 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 7 to destabilize linear RNAs. |
| 262665 | pcDNA3.1(+) ZKSCAN1 Scarless-Split eGFP (mascRNA Mut 10) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using ZKSCAN1 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 10 to destabilize linear RNAs. |
| 262666 | pcDNA3.1(+) Laccase2 Scarless-Split eGFP (mascRNA WT) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using Laccase2 introns. Instead of a poly(A) signal, the primary transcript ends in WT mascRNA to destabilize linear RNAs. |
| 262667 | pcDNA3.1(+) Laccase2 Scarless-Split eGFP (mascRNA Mut 7) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using Laccase2 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 7 to destabilize linear RNAs. |
| 262668 | pcDNA3.1(+) Laccase2 Scarless-Split eGFP (mascRNA Mut 10) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using Laccase2 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut10 to destabilize linear RNAs. |
| 262669 | pcDNA3.1(+) POLR2A Scarless-Split eGFP (mascRNA WT) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using POLR2A introns. Instead of a poly(A) signal, the primary transcript ends in WT mascRNA to destabilize linear RNAs. |
| 262670 | pcDNA3.1(+) POLR2A Scarless-Split eGFP (mascRNA Mut 7) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using POLR2A introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 7 to destabilize linear RNAs. |
| 262671 | pcDNA3.1(+) POLR2A Scarless-Split eGFP (mascRNA Mut 10) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using POLR2A introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 10 to destabilize linear RNAs. |
| 262672 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (mascRNA WT) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns. Instead of a poly(A) signal, the primary transcript ends in WT mascRNA to destabilize linear RNAs. |
| 262673 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (mascRNA Mut 7) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 7 to destabilize linear RNAs. |
| 262674 | pcDNA3.1(+) IGF2BP1 Scarless-Split eGFP (mascRNA Mut 10) | Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using IGF2BP1 introns. Instead of a poly(A) signal, the primary transcript ends in mascRNA Mut 10 to destabilize linear RNAs. |
| 262675 | pCIRCUS dTomato + WT mascRNA + ZKSCAN1 Scarless-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA, wildtype mouse mascRNA, and a circRNA encoding eGFP. ZKSCAN1 introns drive circRNA production. |
| 262676 | pCIRCUS dTomato + WT mascRNA + Laccase2 Scarless-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA, wildtype mouse mascRNA, and a circRNA encoding eGFP. Laccase2 introns drive circRNA production. |
| 262677 | pCIRCUS dTomato + WT mascRNA + Tornado-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA, wildtype mouse mascRNA, and a circRNA encoding scarred eGFP. The Tornado platform drives circRNA production. |
| 262678 | pCIRCUS dTomato + mascRNA Mut 7 + ZKSCAN1 Scarless-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA and a circRNA encoding eGFP. ZKSCAN1 introns drive circRNA production. Mutant mascRNA is post-transcriptionally degraded. |
| 262679 | pCIRCUS dTomato + mascRNA Mut 7 + Laccase2 Scarless-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA and a circRNA encoding eGFP. Laccase2 introns drive circRNA production. Mutant mascRNA is post-transcriptionally degraded. |
| 262680 | pCIRCUS dTomato + mascRNA Mut 7 + Tornado-Split eGFP | CIRCUS platform enabling co-expression of a linear dTomato mRNA and a circRNA encoding eGFP. The Tornado platform drives circRNA production. Mutant mascRNA is post-transcriptionally degraded. |
| 262681 | pCIRCUS dTomato + mascRNA Mut 7 + Tornado-RAB7A Guide | CIRCUS platform enabling co-expression of a linear dTomato mRNA and a circRNA encoding a guide RNA that targets ADAR to RAB7A. The Tornado platform drives circRNA production. |
| 262682 | pCIRCUS dTomato + mascRNA Mut 7 + Tornado-Control Guide | CIRCUS platform enabling co-expression of a linear dTomato mRNA and a circRNA encoding a control guide RNA. The Tornado platform drives circRNA production. |