公开(公告)号：US20220025396A1

公开(公告)日：2022-01-27

申请号：US17052097

申请日：2019-05-07

Applicant: Spark Therapeutics, Inc.

Inventor： Guang QU ,  Denis PHICHITH ,  Jingmin ZHOU

IPC: C12N15/86

Abstract: Disclosed herein are packaging cell lines, in which adenovirus (Ad) E1A is constitutively expressed, that also contain integrated AAV rep and cap genes. The packaging cell lines exhibit little to no expressed Rep protein until helper virus function, such as adenovirus (Ad) E4, E2A and/or VA RNA are provided by, for example, transduction of the cells with a virus, vector or plasmid, such as an Ad-AAV hybrid virus. The promoter driving expression of AAV rep gene can be positioned far enough upstream (5′) of the rep coding sequence that E1A is unable to activate the promoter, activate substantial transcription of the rep gene and in turn produce Rep protein. Introduction of helper virus function, such as E2A, E4 and/or VA RNA into these packaging cells is able to drive AAV rep gene transcription, subsequent Rep protein expression and production of rAAV vector particles.

公开(公告)号：US20190078099A1

公开(公告)日：2019-03-14

申请号：US16088693

申请日：2017-03-30

Applicant: SPARK THERAPEUTICS, INC.

Inventor： Jingmin ZHOU ,  Guang QU ,  John Fraser WRIGHT

IPC: C12N15/52 ,  C07K14/705 ,  C12N15/85 ,  C12N9/06 ,  C12N9/22 ,  C12N9/00 ,  C12N9/12 ,  C12N15/62

Abstract: Cells and cell lines are disclosed that are able to produce therapeutic proteins, antibodies, vectors, and viral vectors such as lentiviral vectors and adeno-associated viral (AAV) vectors. The cells and/or cell lines can have mutations or deletions in either one or both of the endogenous di-hydrofolate reductase (DHFR−/−) or glutamine synthetase (GS−/−) genes such that DHFR and/or GS expression or function is substantially reduced or eliminated.