Multi-target editing vector and its construction method and application

A construction method and multi-target technology, applied in the field of gene editing, can solve the problems of cumbersome experimental operation steps, limited gene editing efficiency, easy to produce off-target effects, etc., to expand the scope of editing, simplify experimental design steps, and reduce experimental costs Effect
CN109136248BActive Publication Date: 2022-02-01GENEWIZ INC SZ
2 Cites 0 Cited by

Patent Information

Authority / Receiving Office
CN ยท China
Patent Type
Patents(China)
Current Assignee / Owner
GENEWIZ INC SZ
Publication Date
2022-02-01

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The invention discloses a multi-target editing vector, which comprises a Cpfl protein expression element and at least two sgRNA transcription elements. The multi-target editing vector realizes the simultaneous editing of different target sites of the genome by co-expressing Cpfl protein and multiple sgRNAs, improves the efficiency of gene editing, reduces the off-target effect of gene editing, and improves the accuracy of gene editing. The present invention also discloses a method for constructing the above-mentioned multi-target editing vector. Through two-step Golden Gate cloning, multiple sgRNAs are serially connected to the second vector with Cpfl protein expression element and fluorescent expression element to obtain the above-mentioned Multi-target editing vectors. The above-mentioned construction method has simple steps, simplifies the experimental design process, and reduces the cost of gene editing. The invention also discloses a kit including the above-mentioned multi-target site carrier, which can be used for co-editing of multi-target sites with high efficiency and low off-target.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention belongs to the technical field of gene editing, and in particular relates to a multi-target editing vector and its construction method and application. Background technique

[0002] Gene editing technology is a technology that can precisely modify the genome or transcripts, and can complete gene site-directed mutation, knockout or knockin of fragments, etc. In the post-genome era, gene editing technology has become an important research content in the field of life sciences. The traditional gene editing technology is based on embryonic stem cells and gene recombination for targeted modification of biological genomes, but this technology has disadvantages such as low targeting efficiency, long experimental cycle and narrow application range. With the continuous development of gene editing technology, artificial nuclease-mediated gene editing technology has begun to be widely used. This technology specifically recognizes and cleaves the ta...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More