公开(公告)号：US20220372497A1

公开(公告)日：2022-11-24

申请号：US17323603

申请日：2021-05-18

Applicant: SHANGHAITECH UNIVERSITY

Inventor： Yajing LIU ,  Shisheng HUANG ,  Xingxu HUANG

IPC: C12N15/62 ,  C12N9/22 ,  C07K14/775

Abstract: The present disclosure relates to the field of biotechnology, in particular to a base editing tool and use thereof. The present disclosure provides a fusion protein comprising a first nCas9 fragment, a chimeric insertion fragment, a second nCas9 fragment and two UGI fragments from N-terminus to C-terminus, wherein the chimeric insertion fragment is selected from APOBEC1 fragment or APOBEC3A fragment. The present disclosure provides a novel base editing tool that is compatible with insertion of various deaminases on the chimeric sites of nCas9. Compared with nCas9 terminal fusion base editor, the base editing tool of the present invention significantly reduce of off-targeting on both DNA and RNA, while maintaining specific targeted base editing efficiency, with higher specificity and favorable industrialization prospects.

公开(公告)号：US20230313205A1

公开(公告)日：2023-10-05

申请号：US18150778

申请日：2023-01-05

Applicant: ShanghaiTech University

Inventor： Yajing LIU ,  Shisheng HUANG ,  Xingxu HUANG

IPC: C12N15/62 ,  C07K14/775 ,  C12N9/22

CPC classification number: C12N15/625 ,  C07K14/775 ,  C12N9/22 ,  C07K2319/09

Abstract: A fusion protein which may comprise a first nCas9 fragment, a chimeric insertion fragment, a second nCas9 fragment and two UGI fragments from N-terminus to C-terminus, wherein the chimeric insertion fragment is selected from APOBEC1 fragment or APOBEC3A fragment for cytosine deamination at the target site. The fusion protein may comprise a first nCas9 fragment, a chimeric insertion fragment and a second nCas9 fragment from N-terminus to C-terminus, wherein the chimeric insertion fragment is TadA-TadA* for cytosine deamination at the target site. The present disclosure provides a novel base editing tool that is compatible with insertion of various deaminases on the chimeric sites of nCas9. Compared with nCas9 terminal fusion base editor, the base editing tool of the present invention significantly reduce off-targeting on both DNA and RNA, while maintaining specific targeted base editing efficiency, with higher specificity and favorable industrialization prospects.