US 12,247,196 B2
Engineered microorganisms and methods of making and using same
Gautam Dantas, St. Louis, MO (US); Suryang Kwak, St. Louis, MO (US); and Herbert Virgin, St. Louis, MO (US)
Assigned to Washington University, St. Louis, MO (US)
Appl. No. 17/278,027
Filed by Washington University, St. Louis, MO (US); and Vir Biotechnology, Inc., San Francisco, CA (US)
PCT Filed Sep. 20, 2019, PCT No. PCT/US2019/052044
§ 371(c)(1), (2) Date Mar. 19, 2021,
PCT Pub. No. WO2020/061389, PCT Pub. Date Mar. 26, 2020.
Claims priority of provisional application 62/733,896, filed on Sep. 20, 2018.
Prior Publication US 2022/0033764 A1, Feb. 3, 2022
Int. Cl. C12N 1/20 (2006.01); C12N 9/22 (2006.01); C12N 15/63 (2006.01); C12R 1/865 (2006.01)
CPC C12N 1/20 (2013.01) [C12N 9/22 (2013.01); C12N 15/63 (2013.01); C12R 2001/865 (2021.05)] 27 Claims
OG exemplary drawing
 
1. An engineered Saccharomyces boulardii cell comprising a genome-integrated display cassette encoding:
a. a signal peptide;
b. a binding agent with binding affinity to a target microorganism selected from the group consisting of norovirus, Clostridium, Listeria, Streptococcus, Staphylococcus, and Campylobacter;
c. an anchor comprising a cell wall anchor protein, or a fragment thereof selected from the group consisting of Cwp2p, Sed1p, Ccw12p, Flo1p and Flo428;
d, wherein (a) is operably linked to (b) which is operably linked to (c);
wherein the genome integrated display cassette is integrated at any one of a site selected from the group consisting of IS1n, IS2n, IS3n, IS4n, IS5n, and IS6n in the Saccharomyces boulardii genome; and
wherein the target microorganism is norovirus and the binding agent is CLM-1; the target microorganism is Clostridium difficile or Clostridium perfringens and the binding agent is a cell binding domain of ΦCP26F; the target microorganism is Listeria monocytogenes and the binding agent is cell binding domain 500 of endolysin Ply500 or cell wall binding domain of PlyP35; the target microorganism is Streptococcus pyogenes and the binding agent is PlyC binding domain of endolysin PlyC; the target microorganism is methicillin-resistant Staphylococcus aureus (MRSA) or Staphylococcus aureus and the binding agent is LysGH15B, a cell wall binding domain of endolysin LysGH15, LysSA97, or ZW88; or the target microorganism is Campylobacter coli and the binding agent is a cell wall binding domain of phiCcoIBB35.