Method for detecting off-target effect of adenine base editor system based on whole genome sequencing, and its application in gene editing

A whole-genome sequencing and editing system technology, applied in the field of detection of off-target effects of adenine single base editing system, can solve problems restricting the application of ABE system and achieve the effect of wide application prospects

Pending Publication Date: 2019-02-01
SUN YAT SEN UNIV
View PDF1 Cites 22 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no method that can detect off-target effects of the ABE system on a genome-wide scale, which seriously restricts the application of the ABE system

Method used

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
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting off-target effect of adenine base editor system based on whole genome sequencing, and its application in gene editing
  • Method for detecting off-target effect of adenine base editor system based on whole genome sequencing, and its application in gene editing
  • Method for detecting off-target effect of adenine base editor system based on whole genome sequencing, and its application in gene editing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0048] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any form.

[0049] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field. Unless otherwise specified, the reagents and materials used in the following examples are commercially available. Experimental methods that do not indicate specific conditions are usually implemented under conventional conditions or conditions suggested by the manufacturer.

[0050] In a specific embodiment of the present invention, the present invention provides a system, method, kit and application thereof for detecting off-target effects of an adenine single base editing system based on whole genome sequencing.

[0051] The method for detecting the off-target effect of the adenine single base editing sy...

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

PUM

No PUM Login to view more

Abstract

The invention provides a method for detecting the off-target effect of an adenine base editor system based on whole genome sequencing, and its application in gene editing, The adenine base editor system comprises a TadA:TadA*:Cas9 fusion gene and gRNA. The system can catalyze the efficient replacement of adenine (A) at a target site with guanine (G), and has a broad application prospect in human disease gene editing therapy and disease model construction. The first method for detecting the off-target effect of the ABE system in a whole genome range, called EndoV-seq method for short, is developed in the invention. The EndoV-seq method provided by the invention has a broad application prospect in the field of gene editing, especially gene editing therapy.

Description

technical field [0001] The invention belongs to the technical field of molecular biology. More specifically, it relates to a method for detecting off-target effects of an adenine base editor (ABE) system based on whole genome sequencing and its application in gene editing. Background technique [0002] CRISPR / Cas9 system is a new artificial nuclease technology, which is a complex composed of gRNA (guide RNA) and Cas9 protein. With the help of the PAM (Protospacer adjacent motif) sequence at the 3' end of the target site, the gRNA-Cas9 protein complex binds to the target DNA through the 20 bases at the 5' end of the gRNA, thereby recruiting the endonuclease Cas9 to the target site point, the target DNA is cleaved, thereby editing the target gene. Although the emergence of CRISPR / Cas9 technology has greatly improved the efficiency of gene site-directed mutation, it still cannot meet the needs of clinical gene therapy. Recently, based on CRISPR / Cas9 technology, scientists ha...

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

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2521/301C12Q2521/539C12Q2535/122
Inventor 松阳洲梁普平黄军就
Owner SUN YAT SEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap