US 11,732,274 B2
Methods and compositions for evolving base editors using phage-assisted continuous evolution (PACE)
David R. Liu, Lexington, MA (US); Ben Thuronyi, Williamstown, MA (US); and Christopher Gerard Wilson, Waltham, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US); and The Broad Institute, Inc., Cambridge, MA (US)
Appl. No. 16/634,405
Filed by President and Fellows of Harvard College, Cambridge, MA (US); and The Broad Institute, Inc., Cambridge, MA (US)
PCT Filed Jul. 27, 2018, PCT No. PCT/US2018/044242
§ 371(c)(1), (2) Date Jan. 27, 2020,
PCT Pub. No. WO2019/023680, PCT Pub. Date Jan. 31, 2019.
Claims priority of provisional application 62/538,380, filed on Jul. 28, 2017.
Prior Publication US 2020/0172931 A1, Jun. 4, 2020
Int. Cl. C12N 15/86 (2006.01); C12N 9/22 (2006.01); C12N 15/113 (2010.01); C12N 9/24 (2006.01); C12N 9/78 (2006.01); C07K 14/32 (2006.01)
CPC C12N 15/86 (2013.01) [C12N 9/22 (2013.01); C12N 15/113 (2013.01); C07K 2319/80 (2013.01); C12Y 305/04005 (2013.01)] 32 Claims
 
1. A cytidine deaminase comprising an amino acid sequence that is at least 90% identical to amino acid residues 3-229 of SEQ ID NO: 2, wherein the cytidine deaminase comprises one or more mutations selected from the group consisting of E4X1, V10X2, E31X3, Y40X4, E95X5, H109X6, H122X7, D124X8, R154X10, N158X11, A165X12, P201X13, F205X14, and I208X15 relative to SEQ ID NO: 2, wherein X1, X3, and X5 are any amino acid other than E, X2 is any amino acid other than V, X4 is any amino acid other than Y, X6 is N, X7 is any amino acid other than H, X8 is any amino acid other than D, X10 is any amino acid other than R, X11 is any amino acid other than N, X12 is any amino acid other than A, X13 is any amino acid other than P, X14 is any amino acid other than F, and X15 is any amino acid other than I.