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Construction method of recombinant sendai virus and application

A Sendai virus and construction method technology, applied in the direction of virus, virus/bacteriophage, recombinant DNA technology, etc., can solve the problems of low reprogramming efficiency, packaging virus purity and easy introduction of other active viruses, etc., to improve packaging efficiency and excellent virus packaging effect, the effect of enhancing the effect of virus expression

Active Publication Date: 2021-03-23
梅尔顿(深圳)生物医药技术有限公司
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for constructing recombinant Sendai virus, which aims to solve the problems of low reprogramming efficiency and easy introduction of other active viruses in the method of cell reprogramming to induce iPS cells

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  • Construction method of recombinant sendai virus and application
  • Construction method of recombinant sendai virus and application
  • Construction method of recombinant sendai virus and application

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Embodiment Construction

[0041] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0042] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships, for example, A and / or B may mean: A exists alone, A and B exist simultaneously, and B exists alone Condition. Among them, A and B can be singular or plural. The character " / " generally indicates that the contextual objects are an "or" relationship.

[0043] In this application, "at least one" means one or more, and "multiple" means two or more. "At least one of the following" or similar expressions refer to any combination of these ...

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Abstract

The invention provides a construction method of a recombinant sendai virus. The method comprises the following steps of constructing plasmids pCAGGS-NP / P / L; amplifying SeV-01 / 02 DNA by taking a sendaivirus reverse transcription sample as a template; amplifying linearized pUC57-insert plasmids, and collecting pUC-insert DNA; fusing SeV-01 / 02 DNA fragments with the pUC-insert DNA to obtain Sev-fullplasmids, and carrying out enzyme digestion to obtain Sev-full-Not1; synthesizing pUC57-A plasmids, carrying out enzyme digestion to collect A-Not1, and connecting the A-Not1 with the Sev-full-Not1 to obtain Sev-A plasmids; and providing 293T cells, adding three kinds of plasmids of pCAGGS-NP / P / L, Sev-A and T7-opt, and carrying out transfection to obtain the recombinant sendai virus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing a recombinant Sendai virus, a kit for reprogramming PBMCs into iPS cells, and a method for reprogramming PBMCs into iPS cells. Background technique [0002] Sendai virus (SeV), also known as type 1 parainfluenza virus or HVJ (Hemagglutinating Virus of Japan, Japanese hemagglutination virus), is a negative-sense single-stranded single-stranded RNA virus of the Paramyxoviridae family. Sendai virus has the following characteristics: (1) During the life cycle of Sendai virus, the viral protein and genome are in the cytoplasm, and the viral genome does not integrate with the cell genome; (2) Sendai virus can infect cells throughout their life cycle and can Infect most cells with extremely high efficiency, including cell types that are difficult to infect with lentivirus; (3) The expression of foreign genes is highly efficient and regulated, and highly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N7/01
CPCC12N15/85C12N7/00C12N2800/107C12N2760/18852
Inventor 张曦萧哲
Owner 梅尔顿(深圳)生物医药技术有限公司
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