Jeremy Wilusz Lab: Engineering circular RNA expression systems to minimize contaminating linear RNA byproducts

Unpublished

Plasmids from Article

ID	Plasmid	Purpose
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
262623	pcDNA3.1(+) POLR2A Scarless-Split eGFP	Expresses an eGFP circular RNA containing the EMCV IRES in mammalian cells using POLR2A introns
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
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
262637	pcDNA3.1(+) circRNA Mini Scarless-ciRS-7	Expresses scarless ciRS-7 (CDR1as) circRNA in mammalian cells using mini 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
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
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
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
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.
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.
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.
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.
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.