Patent ID: 11891609
Assignee: INSCRIPTA, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 10:
11. The method of claim 1, wherein the method further comprises:
(d) providing to the edited cells:
a second editing vector comprising the second inducible promoter or the constitutive promoter driving expression of a coding sequence for a recA protein, wherein the second inducible promoter or the constitutive promoter drives transcription of at least two editing cassettes, wherein each editing cassette comprises:
(A) a gRNA sequence and a donor DNA sequence to be transcribed;
(B) a bacterial origin or replication; and
(C) a third selection marker;

(e) allowing transcription of the recA protein and the at least two editing cassettes from the second inducible promoter or the constitutive promoter; and
(f) following transcription of the recA protein and the at least two editing cassettes, inducing transcription of the CRISPR nuclease to produce twice edited cells.