Exosome delivery CasRx gene silencing AAV vector and construction method and application thereof

A gene silencing and exosome technology, applied in the field of genes to eliminate off-target effects and small size
CN113025656AInactive Publication Date: 2021-06-25THE SECOND PEOPLES HOSPITAL OF SHENZHEN
2 Cites 2 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
Publication Date
2021-06-25
Estimated Expiration
Not applicable · inactive patent

Smart Images

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

Abstract

The invention discloses an exosome delivery CasRx gene silencing AAV vector. The AAV vector contains CasRx protein expression, and a U6 promoter drives sgRNA expression. The invention discloses a construction method of the exosome delivery CasRx gene silencing AAV vector. According to the construction method, when the AAV vector is separated from a conditioned medium for producing cells, AAV can be combined with exosome exoAAV, and efficient delivery of the gene vector can be realized by wrapping the AAV vector with the exosome. Compared with the prior art, the AAV vector has the beneficial effects that 1, the gene knockout efficiency is higher (greater than 90%); 2, compared with RNA interference (shRNA), the off-target effect is eliminated; 3, the AAV vector has the capability of targeting nuclear RNA (including non-coding RNA); 4, splicing is manipulated, wherein misconnection defects and targeted exon jumping are corrected; and 5, the size is small, a single vector AAV-CasRx can be realized, and RNA can be guided in vivo.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention relates to the field of gene technology, in particular to an AAV vector for delivery of CasRx gene silencing by exosomes and its construction method and application. Background technique

[0002] On March 16, 2018, a major achievement published in the journal "Cell" brought a big leap forward in RNA editing technology. Scientists from the Salk Institute in the United States expanded the RNA editing ability with a new CRISPR family enzyme, and Name this new system "CasRx." A key feature of the new system, in addition to its high efficiency and absence of apparent off-target effects, is its reliance on an enzyme that is physically smaller than in previous studies. This is crucial for RNA editing technology, which makes it easier for the editing tool to be packaged into a viral vector and enter cells for RNA editing. Hiroshi Nishimasu, a scientist at the University of Tokyo who was not involved in the study, said: "In this study, the resea...

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