SG11201907273RA - Rebooting of synthetic bacteriophage genome in l-form bacteria - Google Patents

Abstract

INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 16 August 2018 (16.08.2018) WIP0 1 PCT ill mu °million °nolo oimIE (10) International Publication Number WO 2018/146206 Al (51) International Patent Classification: C07K 14/195 (2006.01) Cl 2N 7/00 (2006.01) (21) International Application Number: PCT/EP2018/053202 (22) International Filing Date: 08 February 2018 (08.02.2018) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 17155693.9 10 February 2017 (10.02.2017) EP (71) Applicant: ETH ZURICH [CH/CH]; Raemistrasse 101 / ETH transfer, 8092 Zurich (CH). (72) Inventors: LOESSNER, Martin; Stuhlenstrasse 39, 8123 Ebmatingen (CH). KILCHER, Samuel; Nidelbadstrasse 43, 8803 Riischlikon (CH). STUDER, Patrick; Winzer- strasse 62, 8049 Ziirich (CH). (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, JO, 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, (54) Title: REBOOTING OF SYNTHETIC BACTERIOPHAGE GENOME IN L-FORM BACTERIA Fig. 1 A L-form Transformation and + genomic DNA phage re b oo ti ng + PEG INDICATOR STRAIN mix L-forms and propagation strain at propagation strain O 00 O O (57) : The present invention related to a method for producing or propagating an engineered bacteriophage, comprising the steps of: providing a functional synthetic genome of an engineered bacteriophage being able to infect a target bacterium; providing a recipient bacterium; transforming the recipient bacterium with the functional synthetic genome in a transformation step, yielding a transformed recipient bacterium; incubating the transformed recipient bacterium in a first incubation step, wherein the engineered bacteriophage is propagated within the transformed recipient bacterium; and further incubating the transformed recipient bacterium or the propagated engineered bacteriophage released from the transformed recipient bacterium with the target bacterium in a second incubation step, wherein the propagated engineered bacteriophage infects the target bacterium and is further propagated within the target bacterium, and wherein the recipient bacterium is a cell wall-deficient bacterium. [Continued on next page] WO 2018/146206 Al MIDEDIMOMMIDIREEMOMMIERIDIONEONVOIMIE 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))