Cpf1-based DNA in-vitro splicing method

A technology of motif and reaction system, applied in the field of DNA splicing in vitro based on Cpf1
CN107881184AActive Publication Date: 2018-04-06CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
Publication Date
2018-04-06

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Abstract

The invention relates to a Cpf1-based DNA in-vitro splicing method. According to the method, a specific crRNA sequence capable of being recognized by CRISPR-Cpf1 is designed and synthesized and is utilized for guiding Cpf1 to specifically cutting off a specific position of double-stranded DNA and generate preset cohesive ends. By virtue of the method, the preset cohesive ends can be conveniently,rapidly and accurately acquired and are utilized for splicing DNA; and engineering, standardized and modularized remolding or assembling of DNA can be realized.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a Cpf1-based DNA splicing method in vitro. The method of the invention can generate predetermined sticky ends, and can be used for DNA seamless splicing in vitro. Background technique

[0002] In 2010, J. Craig Venter's laboratory completed the artificial construction of Mycoplasma mycoides "Synthia", setting off a storm of synthetic biology research. Compared with the traditional restriction endonuclease digestion-ligase ligation cloning method, seamless splicing has many advantages such as convenient design and good compatibility because it does not introduce additional sequences, and it plays an increasingly important role in the development of synthetic biology. role.

[0003] The existing seamless splicing technologies mainly include splicing methods based on Type IIS restriction endonucleases, splicing methods based on specific base modifications, and s...

Claims

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