公开(公告)号：US10725041B2

公开(公告)日：2020-07-28

申请号：US14931321

申请日：2015-11-03

Applicant: Versiti Blood Research Institute Foundation, Inc.

Inventor： Peter Jay Newman ,  Sridhar Rao ,  Nanyan Zhang ,  Huiying Zhi

IPC: G01N33/569 ,  C12N15/85 ,  C12N5/078 ,  C12N5/074 ,  C12N15/11

Abstract: Methods of creating cells expressing specific platelet alloantigens by combining gene editing techniques and cell culture differentiation or expansion techniques employing pluripotent cells, including the steps of transfecting pluripotent cells with a plasmid encoding one or more guide RNAs targeting within a platelet alloantigen target locus and a nuclease in the presence of an HDR repair oligo and culturing the resulting cells to expand their numbers or to create a differentiated cell type of interest.

公开(公告)号：US20220211017A1

公开(公告)日：2022-07-07

申请号：US17668086

申请日：2022-02-09

Applicant: VERSITI BLOOD RESEARCH INSTITUTE FOUNDATION, INC.

Inventor： Peter J. Newman ,  Huiying Zhi

IPC: A01K67/027 ,  C07K14/705 ,  G01N33/50 ,  C12N15/89 ,  G01N33/577

Abstract: A transgenic mouse comprising T30A, S32P, Q33L, N39D, and M470Q mutations in GPIIIa, as well as methods for making the transgenic mouse and methods for using the transgenic mouse to screen test compounds are described.

公开(公告)号：US11266129B2

公开(公告)日：2022-03-08

申请号：US16674804

申请日：2019-11-05

Applicant: VERSITI BLOOD RESEARCH INSTITUTE FOUNDATION, INC.

Inventor： Peter J. Newman ,  Huiying Zhi

IPC: G01N33/566 ,  A01K67/027 ,  C07K14/705 ,  G01N33/50 ,  C12N15/89 ,  G01N33/577

Abstract: A transgenic mouse comprising T30A, S32P, Q33L, N39D, and M470Q mutations in GPIIIa, as well as methods for making the transgenic mouse and methods for using the transgenic mouse to screen test compounds are described.

公开(公告)号：US20210127648A1

公开(公告)日：2021-05-06

申请号：US16674804

申请日：2019-11-05

Applicant: VERSITI BLOOD RESEARCH INSTITUTE FOUNDATION, INC.

Inventor： Peter J. Newman ,  Huiying Zhi

IPC: A01K67/027 ,  C07K14/705 ,  G01N33/50 ,  G01N33/577 ,  C12N15/89

Abstract: A transgenic mouse comprising T30A, S32P, Q33L, N39D, and M470Q mutations in GPIIIa, as well as methods for making the transgenic mouse and methods for using the transgenic mouse to screen test compounds are described.

公开(公告)号：US20200332259A1

公开(公告)日：2020-10-22

申请号：US16913741

申请日：2020-06-26

Applicant: VERSITI BLOOD RESEARCH INSTITUTE FOUNDATION, INC.

Inventor： PETER JAY NEWMAN ,  Sridhar Rao ,  Nanyan Zhang ,  Huiying Zhi

IPC: C12N5/0789 ,  C12N9/22 ,  C12N15/113 ,  C12N5/078

Abstract: A method of creating cells expressing specific red blood cell antigens is disclosed. In one embodiment, the method comprises the steps of (a) combining one or more guide RNAs targeting within a red blood cell antigen target locus; (b) adding a repair template comprising a mutation in the target locus flanked by a homology arm on each side, wherein the template may additionally include a diagnostic restriction enzyme site at the target locus; (c) ligating the guide sequence of step (b) into a plasmid which also expresses a nuclease and, optionally, a selectable marker or a reporter gene; (d) transfecting pluripotent cells with the plasmid of step (c) in the presence of an HDR repair oligo; (e) cloning and testing the resulting reporter positive clones for expression of the antigen target of interest; and (f) culturing the resulting cells to expand their numbers or to create a differentiated cell type of interest.