公开(公告)号：US12157907B2

公开(公告)日：2024-12-03

申请号：US18052783

申请日：2022-11-04

Applicant: MODERNA ENZYMATICS CO., LTD.

Inventor： Masayuki Su'etsugu ,  Seia Nara

IPC: C12Q1/68 ,  C12N15/10 ,  C12P19/34

Abstract: Provided is a method capable of replicating or amplifying circular DNA, and particularly, long-chain circular DNA, in a cell-free system. Specifically, provided is a method for suppressing generation of a DNA multimer as a by-product, when circular DNA having a replication origin sequence (origin of chromosome (oriC)) is replicated or amplified by using the following enzyme groups: (1) a first enzyme group that catalyzes replication of circular DNA; (2) a second enzyme group that catalyzes an Okazaki fragment maturation and synthesizes two sister circular DNAs constituting a catenane; and (3) a third enzyme group that catalyzes a separation of two sister circular DNAs. Moreover, also provided is a method comprising introducing oriC into circular DNA by using a transposon.

公开(公告)号：US20240018561A1

公开(公告)日：2024-01-18

申请号：US18052783

申请日：2022-11-04

Applicant: MODERNA ENZYMATICS CO., LTD.

Inventor： Masayuki Su'etsugu ,  Seia Nara

IPC: C12P19/34 ,  C12N15/10

CPC classification number: C12P19/34 ,  C12N15/10

Abstract: Provided is a method capable of replicating or amplifying circular DNA, and particularly, long-chain circular DNA, in a cell-free system. Specifically, provided is a method for suppressing generation of a DNA multimer as a by-product, when circular DNA having a replication origin sequence (origin of chromosome (oriC)) is replicated or amplified by using the following enzyme groups:
(1) a first enzyme group that catalyzes replication of circular DNA;
(2) a second enzyme group that catalyzes an Okazaki fragment maturation and synthesizes two sister circular DNAs constituting a catenane; and
(3) a third enzyme group that catalyzes a separation of two sister circular DNAs. Moreover, also provided is a method comprising introducing oriC into circular DNA by using a transposon.

公开(公告)号：US20250051817A1

公开(公告)日：2025-02-13

申请号：US18920933

申请日：2024-10-20

Applicant: Moderna Enzymatics Co., Ltd.

Inventor： Masayuki Suetsugu ,  Seia Nara

IPC: C12P19/34 ,  C12N15/10

Abstract: Provided is a method capable of replicating or amplifying circular DNA, and particularly, long-chain circular DNA, in a cell-free system. Specifically, provided is a method for suppressing generation of a DNA multimer as a by-product, when circular DNA having a replication origin sequence (origin of chromosome (oriC)) is replicated or amplified by using the following enzyme groups: (1) a first enzyme group that catalyzes replication of circular DNA; (2) a second enzyme group that catalyzes an Okazaki fragment maturation and synthesizes two sister circular DNAs constituting a catenane; and (3) a third enzyme group that catalyzes a separation of two sister circular DNAs. Moreover, also provided is a method comprising introducing oriC into circular DNA by using a transposon.

公开(公告)号：US20240352495A1

公开(公告)日：2024-10-24

申请号：US18685259

申请日：2022-09-13

Applicant: Moderna Enzymatics Co., Ltd.

Inventor： Masayuki Suetsugu ,  Seia Nara

IPC: C12P19/34 ,  C12N9/12 ,  C12N15/11

CPC classification number: C12P19/34 ,  C12N9/1205 ,  C12N9/1217 ,  C12N9/1223 ,  C12N9/1229 ,  C12N15/11 ,  C12Y207/0104 ,  C12Y207/02001 ,  C12Y207/03002 ,  C12Y207/04001 ,  C12Y207/04006

Abstract: Provided is a method for producing circular DNA in which a region that is sandwiched by a region Ha and a region Hb in circular double-stranded DNA is substituted with the entirety or a portion of a linear-DNA fragment, wherein the region Hb is located downstream of the region Ha in the circular double-stranded DNA; the linear-DNA fragment is single-stranded or double-stranded linear DNA that has a homologous region that corresponds to the region Ha and a homologous region that corresponds to the region Hb, the latter homologous region being positioned downstream of the former homologous region; and the method comprising: preparing a reaction solution that contains the circular double-stranded DNA, the linear-DNA fragment, and a protein that has RecA-family-recombinase activity, and performing homologous recombination reaction by incubating the reaction solution for a predetermined period of time, thereby producing circular DNA in which the region that is from the region Ha to the region Hb in the circular double-stranded DNA is substituted with the region that is from the homologous region that corresponds to the region Ha to the homologous region that corresponds to the region Hb in the linear-DNA fragment.