US 12,410,418 B2
Optimized engineered meganucleases having specificity for a recognition sequence in the Hepatitis B virus genome
Janel Lape, Wake Forest, NC (US); and James Jefferson Smith, Morrisville, NC (US)
Assigned to Precision BioSciences, Inc., Durham, NC (US)
Appl. No. 17/782,610
Filed by Precision BioSciences, Inc., Durham, NC (US)
PCT Filed Dec. 4, 2020, PCT No. PCT/US2020/063479
§ 371(c)(1), (2) Date Jun. 3, 2022,
PCT Pub. No. WO2021/113765, PCT Pub. Date Jun. 10, 2021.
Claims priority of provisional application 62/944,862, filed on Dec. 6, 2019.
Prior Publication US 2023/0031465 A1, Feb. 2, 2023
Int. Cl. C12N 9/22 (2006.01); A61P 31/20 (2006.01); C12N 15/10 (2006.01); C12N 15/90 (2006.01)
CPC C12N 9/22 (2013.01) [A61P 31/20 (2018.01); C12N 15/102 (2013.01); C12N 15/90 (2013.01); C12N 2730/10121 (2013.01)] 12 Claims
 
1. An engineered meganuclease that binds and cleaves a recognition sequence consisting of SEQ ID NO: 10 within a Hepatitis B virus genome, wherein said engineered meganuclease comprises a first subunit, a second subunit, and a linker that covalently joins said first subunit and said second subunit, wherein said first subunit binds to a first recognition half-site of said recognition sequence and comprises a first hypervariable (HVR1) region, wherein said HVR1 region comprises residues corresponding to residues 215, 217, 219, 221, 223, 224, 229, 231, 233, 235, 237, 259, 261, 262, 263, 264, 266, and 268 of SEQ ID NO: 12, and wherein said second subunit binds to a second recognition half-site of said recognition sequence and comprises a second hypervariable (HVR2) region, wherein said HVR2 region comprises residues corresponding to residues 24, 26, 28, 30, 32, 33, 38, 40, 42, 44, 46, 51, 68, 70, 75, and 77 of SEQ ID NO: 12,
wherein said first subunit has at least 97% sequence identity to residues 198-344 of SEQ ID NO: 12, and
wherein said second subunit has at least 97% sequence identity to residues 7-153 of SEQ ID NO: 12.