US 12,454,705 B2
Differential knockout of an allele of a heterozygous apolipoprotein A1 (APO1A) gene
David Baram, Tel Aviv (IL); Lior Izhar, Tel Aviv (IL); Asael Herman, Ness Ziona (IL); Rafi Emmanuel, Ramla (IL); Michal Golan Mashiach, Ness Ziona (IL); and Joseph Georgeson, Rehovot (IL)
Assigned to EMENDOBIO INC., Wilmington, DE (US)
Appl. No. 17/041,661
Filed by EMENDOBIO INC., Wilmington, DE (US)
PCT Filed Mar. 22, 2019, PCT No. PCT/US2019/023715
§ 371(c)(1), (2) Date Sep. 25, 2020,
PCT Pub. No. WO2019/190930, PCT Pub. Date Oct. 3, 2019.
Claims priority of provisional application 62/647,745, filed on Mar. 25, 2018.
Prior Publication US 2021/0032663 A1, Feb. 4, 2021
Int. Cl. C12N 15/90 (2006.01); A61K 35/13 (2015.01); C12N 15/11 (2006.01)
CPC C12N 15/907 (2013.01) [A61K 35/13 (2013.01); C12N 15/11 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 14 Claims
 
1. A method for inactivating a mutant allele of the Apolipoprotein A1 (APOA1), the method comprising:
(a) delivering to an isolated human cell that comprises a mutant APOA1 allele and a functional APOA1 allele a composition comprising:
a CRISPR nuclease, and
an isolated guide RNA molecule (gRNA) that targets the mutant human APOA1 allele, wherein the gRNA comprises a CRISPR RNA (crRNA) comprising a nucleic acid sequence of 17-20 nucleotides which comprise 17-20 contiguous nucleotides set forth in SEQ ID NOs: 114, 188, 192, or 253; and
(b) culturing the cell obtained in step a) such that the mutant APOA1 allele is inactivated and the functional APOA1 allele remains intact.