CRISPR single-base restoration system and application thereof

A repair system, single-base technology, applied in the field of gene editing, which can solve problems such as off-target, reduced stability and melting temperature, and slight toxicity
CN109517845APending Publication Date: 2019-03-26THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
3 Cites 8 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
Publication Date
2019-03-26

Smart Images

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

Abstract

The invention discloses a CRISPR single-base restoration system. The CRISPR single-base restoration system is capable of expressing eukaryotic expression vectors of sgRNA and Cas9 and single strandedoligonucleotide as a donor template, wherein the base sequence of single stranded oligonucleotide is basically the same as the base sequence between bases 30-60 on two sides of a to-be-restored singlebase, and the differences between the two base sequences are that the base corresponding to the to-be-restored single base is a non-mutation wild base, and nonsense mutation occurs on one side of theto-be-restored single base. The invention further discloses application of the CRISPR single-base restoration system in the preparation of drugs for treating beta thalassemia caused due to CD17A-T point mutation of a beta bead protein gene. According to the CRISPR single-base restoration system, a CRISPR / Cas expression plasmid and a donor are commonly transfected into cells, the traceless directional transformation of gene mutation sites in beta17 thalassemia is specifically induced in a 293TT cell, and the method has very important clinical application values in the execution of gene editingaccurate treatment on diseases based on single-gene point mutation.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention relates to the technical field of gene editing, in particular to a CRISPR single-base repair system and its application in the treatment of β17 thalassemia genetic diseases caused by single gene point mutations. Background technique

[0002] β-thalassemia (β-thalassemia) is a disease caused by a decrease in or failure to synthesize the globin peptide chains that make up hemoglobin HbA (a2β2) due to mutations or deletions in the β-globin gene, resulting in an imbalance in the synthesis of a and β peptide chains. An autosomal recessive genetic disorder. The point mutation of β-thalassemia gene is often the most common, and the CD17(A→T) point mutation of β-globin gene is one of the earliest reported point mutations related to β-thalassemia disease.

[0003] Thalassemia is currently treated with lifelong blood transfusions, yet the vast majority of children die before adulthood despite aggressive treatment. Another treatment option is bone...

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