Production method for genome-edited cells
AssigneeKyushu University, National University Corporation
InventorsAtsushi Suzuki, Masaki Kawamata
Life of this patent
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Abstract
A production method for a cell in which only one allele is genome-edited includes: a step of introducing, into a cell, (A) at least one selected from the group consisting of (a1) a guide RNA in which one or more nucleotide residues are added to a 5′-end of a spacer sequence, (a2) a guide RNA containing a spacer sequence having single-base or multiple-base mismatches with respect to a target sequence, and (a3) an expression vector that can cause the guide RNA of (a1) or (a2) to be expressed, and (B) at least one selected from the group consisting of a Cas protein and an expression vector that can cause the Cas protein to be expressed.
Claims14
A guide RNA comprising a spacer sequence,
- wherein 15 or more nucleotide residues are added to the 5′-end of the spacer sequence, and
- wherein said 15 or more nucleotide residues comprise a polycytosine or a polyguanine including 15 or more nucleotide residues.
A guide RNA, comprising a spacer sequence and a repeat sequence,
- wherein the spacer sequence targets a target region, and the repeat sequence forms base pairs with an anti-repeat sequence derived from a tracrRNA to form a complex with a Cas protein,
- wherein 15 or more nucleotide residues are added to a 5′-end of the spacer sequence, and wherein said 15 or more nucleotide residues comprise a polycytosine or a polyguanine of 15 or more nucleotide residues,
- wherein the guide RNA suppresses cytotoxicity, increases homology directed repair frequency, or increases mono-allelic genome editing frequency in genome editing compared to a guide RNA which does not have a nucleotide residue added to the 5′ end of the spacer sequence.
Dependencies are inferred from the claim text.