Method for realizing efficient and fixed-point transgenosis by using TALEN (Transcription Activator-Like Effector Nuclease) technology

Abstract

The invention relates to a method for realizing efficient and fixed-point transgenosis by using a TALEN (Transcription Activator-Like Effector Nuclease) technology, belonging to the field of molecular biology. The method comprises the following steps of: designing a TALEN target point with a specific sequence in a special position in an organism genome by using the TALEN technology, constructing a TALEN plasmid, and meanwhile, constructing an exogenous gene plasmid; then, co-transfecting the TALEN plasmid and an exogenous gene into recipient cells, and then incising in a to-be-integrated position of a recipient cell genome; and finally, automatically inserting an exogenous gene sequence to the exogenous gene at the incision of the recipient cell genome. The fixed-point integration of the exogenous gene on the genome is realized by using the method; and the method is simple in operation, high in controllability and strong in practicability.

Description

technical field [0001] The invention relates to a high-efficiency fixed-point transgenic method using TALEN technology, which is a kind of transgenic technology that utilizes TALEN technology to cut a specific position of the genome to efficiently and accurately integrate foreign genes into the genome of an organism, belonging to the field of molecular biology. Background technique [0002] In transgenic technology, conventional methods for transferring foreign genes into organisms include gene overexpression (Over-expression) and gene knock-in (Knock in). Among them, although the gene overexpression method is simple, it can only randomly integrate the foreign gene into the genome of the organism, and cannot arbitrarily integrate the foreign gene into the specific position of the genome we want; gene knock-in (Knock in) Although the method can site-specifically integrate the exogenous gene into the specific position of the genome we want, the method is difficult, inefficien...

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Problems solved by technology

[0004] Aiming at the disadvantages in the transgenic technology that exogenous genes are difficult to be fixed-point integrated into the designated position on the genome and the integration efficiency is low and difficult, the present invention proposes a high-efficiency fixed-point transgenic method using TALEN technology. Form an incision to integrate the exogenous gene at the incision, and realize the fixed-point integration of the exogenous gene on the genome. This method is simple to operate, highly controllable, and strong in practicability

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