Patent SG-11201807374W-A (SG)
Title: Rna replicon for versatile and efficient gene expression
Inventor: BEISSERT TIM
IPC: C12N15/85
Filed: 2017-03-13
Abstract: EcoRVSelf Sil izza. nsP I nsP2 I nsP3 I nsP4 5x ATG ATG SL1 SL2 /4 5'UTR I In 3'ur4 PuklUk Template RNA WT-RRS A5ATG-RRS 5'UTR .5 ' TG A5ATG-RRSASGP AAAA AAAA 5'UTR \"Are ATG SL1 SL2/3/4 Transgene 1 Elm Transgene 2 A5ATG-R115- bicistronic SL1 SL /4 A AAAA sP3 nsP3 n3P1 nsP, cisReplicon I Tit T- 1 ASATG-RRS 5'UTR A5ATG SG gjeTransgene ATG (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/162460 Al 28 September 2017 (28.09.2017) WIPO I PCT 11111111111111011101111111111101011111011101111111101011111111111111111111111110111111 W O 20 17 / 16 2460 Al (51) International Patent Classification: C12N 15/86 (2006.01) C12N 5/10 (2006.01) C12N 15/85 (2006.01) (21) International Application Number: PCT/EP2017/055808 (22) International Filing Date: 13 March 2017 (13.03.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: PCT/EP2016/056165 21 March 2016 (21.03.2016) EP (71) Applicants: BIONTECH RNA PHARMACEUTICALS GMBH [DE/DE]; An der Goldgrube 12, 55131 Mainz (DE). TRON - TRANSLATIONALE ONKOLOGIE AN DER UNIVERSITATZSMEDIZIN DER JOHANNES GUTENBERG-UNIVERSITAT MAINZ GEMEIN- NUTZIGE GMBH [DE/DE]; Freiligrathstr. 12, 55131 Mainz (DE). (72) Inventors: BEISSERT, Tim; Edith-Stein-Weg 4, 64521 Gross-Gerau (DE). SAHIN, Ugur; Philipp-von-Za- bern-Platz 1, 55116 Mainz (DE). PERKOVIC, Mario; Falkenweg 1, 60437 Frankfurt (DE). (74) Agent: SCHNAPPAUF, Georg; ZSP Patentanwalte PartG mbB, Hansastrasse 32, 80686 Mfinchen (DE). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Published: — with international search report (Art. 21(3)) [Continued on next page] = (54) Title: RNA REPLICON FOR VERSATILE AND EFFICIENT GENE EXPRESSION cisReplicon WT-RRS Fig 1 (57) : The present invention embraces a RNA replicon that can be replicated by a replicase of alphavirus origin. The RNA replicon comprises sequence elements required for replication by the replicase, but these sequence elements do not encode any pro - tein or fragment thereof, such as an alphavirus non-structural protein or fragment thereof. Thus, in the RNA replicon according to the invention, sequence elements required for replication by the replicase and protein-coding region(s) are uncoupled. According to the present invention the uncoupling is achieved by the removal of at least one initiation codon compared to a native alphavirus gen - omic RNA. In particular, the RNA replicon comprises a 5' replication recognition sequence, wherein the 5' replication recognition sequence is characterized in that it comprises the removal of at least one initiation codon compared to a native alphavirus 5' replica- tion recognition sequence. The replicase of alphavirus origin may be encoded by an open reading frame on the RNA replicon or on a separate RNA molecule. The present invention enables efficient and safe expression of a protein of interest in a cell or organism, but is not associated with undesired production of fragments of alphavirus non-structural protein. Methods of protein production in vitro and in vivo, as well as medical uses, are provided herein. WO 2017/162460 Al 1#1101MO1101DION01013011101010111100110MOVOIMIE — with sequence listing part of description (Rule 5.2(a))