	US 12,435,334 B2
	Differential knockout of an allele of a heterozygous ELANE gene-II
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)
Filed by EmendoBio Inc., Wilmington, DE (US)
Filed on Nov. 5, 2020, as Appl. No. 17/090,804.
Claims priority of provisional application 62/931,641, filed on Nov. 6, 2019.
Prior Publication US 2021/0155929 A1, May 27, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/113 (2010.01); C12N 9/64 (2006.01); C12N 15/90 (2006.01)

CPC C12N 15/113 (2013.01) [C12N 9/6448 (2013.01); C12N 15/90 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)]

7 Claims

1. A method for inactivating in a cell a mutant allele of the elastase, neutrophil expressed gene (ELANE gene) having a mutation associated with severe congenital neutropenia (SCN) or cyclic neutropenia (CyN) and which cell is heterozygous at the polymorphic site rs1683564, the method comprising introducing to the cell a composition comprising:

a Cas9 CRISPR nuclease or a sequence encoding said CRISPR nuclease; and

a first RNA molecule comprising a guide sequence portion having 17-30 nucleotides,

wherein the guide sequence portion of the first RNA molecule comprises 17-30 nucleotides which comprise 17-20 contiguous nucleotides set forth in any one of SEQ ID NOs: 335-354; and

wherein a complex of the CRISPR nuclease and the first RNA molecule affects a double strand break in the mutant allele of the ELANE gene.