Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A chimeric nuclease that specifically recognizes and repairs the beta-globin gene of β-thalassemia

An amino acid and nucleic acid molecule technology, applied in the field of genetic engineering, which can solve problems such as off-target and obstacles

Active Publication Date: 2019-06-11
TSINGHUA UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, ZFN can only recognize a small number of sites at present, and there will be relatively large limitations in application; TALEN technology has the characteristics of easy assembly, fast speed, and strong recognition, but because TALEN contains FokI, it has non-specificity in recognition, resulting in certain off-target; CRISPR / Cas9 system has the characteristics of simple operation, but its binding to nucleic acid is very non-specific, which will be an important obstacle to its application

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
  • A chimeric nuclease that specifically recognizes and repairs the beta-globin gene of β-thalassemia
  • A chimeric nuclease that specifically recognizes and repairs the beta-globin gene of β-thalassemia
  • A chimeric nuclease that specifically recognizes and repairs the beta-globin gene of β-thalassemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Preparation of a chimeric nuclease that specifically recognizes and repairs the beta-thalassemia beta-globin gene

[0038] 1. Transformation of homing endonuclease I-SceI

[0039] The homing endonuclease I-SceI will use a special fluorescent reporter system in human cells to screen the DNA sequence that specifically recognizes the DNA sequence adjacent to the β-thalassemia mutation site ( figure 1 ). From the current structure of I-SceI, it can be found that some amino acids in I-SceI are in direct contact with the DNA sequence. By gradually changing the sequence of bases, and then screening these directly contacted amino acids, and finding the most specific new combination amino acid binding site (see figure 2 ).

[0040] During the recognition process of I-SceI and DNA sequence, ATAAC in the DNA sequence is an important part of protein cleavage. When these bases are changed, the cleavage efficiency will be significantly reduced, which can be screened by...

Embodiment 2

[0055] Embodiment 2, functional verification of chimeric nuclease TALE-I-SceI

[0056] 1. Improving the efficiency of gene targeting in the human genome

[0057] The modified fusion protein has the effect of cutting genomic DNA. In the absence of a donor, the DSB will be repaired by the NEHJ repair method in vivo, and indels or mutations will be generated at the cutting site.

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

Disclosed is chimeric nuclease TALE-ISceI, in which FokI in TALEN is replaced with fusion protein obtained by transformed I-SceI. The chimeric nuclease TALE-ISceI can specifically recognize and repair a β thalassemia β-globin gene and can be used for treating β-thalassemia.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a chimeric nuclease that specifically recognizes and repairs the beta-globin gene of beta thalassemia. Background technique [0002] Thalassemia is a genetic disease with high incidence and wide distribution in China and the world. Normal hematopoietic stem cells have the ability to differentiate into blood cells. In patients with thalassemia, their red blood cells are abnormal, fragile and easy to die, and their oxygen-carrying capacity is also insufficient. The most effective methods for anemia include bone marrow hematopoietic stem cell transplantation (BMT), peripheral blood stem cell transplantation (PB SCT) and cord blood stem cell transplantation (CBSCT). However, lack of donors and rejection have severely hindered the widespread use of this method. [0003] Beta-thalassemia is a type of anemia caused by mutations in the beta globin gene. Normal ...

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 Patents(China)
IPC IPC(8): C12N9/22C12N15/62C12N15/85A61K38/46A61K48/00A61P7/06
CPCA61K38/46A61K48/00C07K14/00C07K19/00C12N9/22C12N15/62C12N15/85
Inventor 纪家葵林建飞
Owner TSINGHUA UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com