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Gene editing nanocapsule, and preparation method and application thereof

A nanocapsule and gene editing technology, applied in the field of gene editing, can solve the problems of poor cell membrane permeability and low stability, achieve good biocompatibility and biosafety, small particle size, and high cell endocytosis efficiency Effect

Active Publication Date: 2021-06-18
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, significant challenges remain in drug delivery
Most proteins such as enzymes, antibodies or transcription factors face the problems of low stability and poor cell membrane permeability due to their fragile tertiary structure and large molecular size

Method used

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  • Gene editing nanocapsule, and preparation method and application thereof
  • Gene editing nanocapsule, and preparation method and application thereof
  • Gene editing nanocapsule, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Embodiment 1 ANC SS Preparation method of (Cas9 / sgRNA) nanocapsules

[0109] Step 1, Cas9 and sgRNA were added to 500 μl 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES buffer) (10mM PH7.4) at a molar ratio of 1:1.2, and incubated at room temperature 5 minutes.

[0110] Specifically, the target gene of the sgRNA is the PLK1 gene, and the sequence of the target site of the sgRNA on the PLK1 gene is shown in SEQ ID NO:1.

[0111] Step 2. Transfer the above system to a 4°C environment, add acrylate polyethylene glycol succinyl carboxymethyl ester and stir for 10 minutes, then add guanidine acrylate and stir for 5 minutes, then add degradable N,N'-bis (acryloyl)cystamine, wherein the molar ratio of acrylate polyethylene glycol succinylcarboxymethyl ester, guanidinoacrylate, and N,N'-bis(acryloyl)cystamine is 1:1:1. Add 3 mg of ammonium persulfate and 3 μL of N, N, N', N'-tetramethylethylenediamine solution to initiate the polymerization reaction immediately. Th...

Embodiment 2

[0117] Embodiment 2 ANC SS Preparation method of (Cas9 / sgRNA) nanocapsules

[0118] The only difference from the above-mentioned Example 1 is that the molar ratio of the Cas nuclease to the guanidinoacrylate is 1:200.

Embodiment 3

[0119] Embodiment 3 ANC SS Preparation method of (Cas9 / sgRNA) nanocapsules

[0120] The only difference from the above-mentioned Example 1 is that the molar ratio of the Cas nuclease to the guanidinoacrylate is 1:250.

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Abstract

The invention provides a gene editing nanocapsule, and a preparation method and application thereof. The gene editing nanocapsule has a core shell structure, wherein an inner core comprise a Cas / sgRNA ribonucleoprotein compound, an outer shell comprises a polymer, the Cas / sgRNA ribonucleoprotein compound has a gene editing function, and the polymer serves as a carrier of the Cas / sgRNA ribonucleoprotein compound and protects the Cas / sgRNA ribonucleoprotein compound. Since the polymer contains a tumor microenvironment sensitive molecule, the nanocapsule can be efficiently released in tumor cells. Furthermore, the surface of the outer shell can be modified with a targeting agent, the nanocapsule can carry out specific targeting on the tumor cells, and the endocytosis efficiency of the nanocapsule can be improved. The gene editing nanocapsule has good biocompatibility and biosecurity, and therefore, the gene editing nanocapsule is expected to be a safe and efficient gene treatment medicine which aims at tumors.

Description

technical field [0001] The invention relates to the technical field of gene editing, in particular to a gene editing nanocapsule and its preparation method and application. Background technique [0002] Malignant tumors are the primary diseases that threaten human life and health. Most of the tumors have an insidious onset, no obvious symptoms in the early stage, and the diagnosis is already in the middle and late stage, and the chance of surgical resection has been lost. At present, chemotherapy is still one of the main means of tumor treatment. However, traditional chemotherapy drugs have strong side effects, poor targeting, and are prone to drug resistance. Gene therapy is playing an increasingly important role in tumor treatment because it can specifically change genetic material to achieve the purpose of treatment, has strong targeting, less toxic and side effects, and permanent treatment. However, there are many defects such as complex operation, low efficiency, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/46A61P35/00C12N9/22C12N15/113C08F283/06C08F222/38C08F220/60B82Y5/00B82Y30/00B82Y40/00
CPCA61K48/0041A61K38/465B82Y5/00B82Y30/00B82Y40/00A61P35/00C12N9/22C12N15/113C08F283/065C12N2310/20C08F222/385C08F220/60A61K47/62A61K47/6925A61K47/58A61K31/7105Y02A50/30C12N15/11C12N2310/351C12N11/04C12N2310/3513
Inventor 邹艳师冰洋孙新红郑蒙
Owner HENAN UNIVERSITY