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Variant Gsdmd-C nucleic acid fragment, carrier containing nucleic acid fragment, and application

A nucleic acid fragment and vector technology, applied in the field of biomedicine

Inactive Publication Date: 2020-10-20
上海朗昇生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, in the prior art, there are no reports about the adeno-associated virus vector expressing Gsdmd-C protein and its preparation method and its application in the treatment of retinal detachment photoreceptor cell injury and death.

Method used

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  • Variant Gsdmd-C nucleic acid fragment, carrier containing nucleic acid fragment, and application
  • Variant Gsdmd-C nucleic acid fragment, carrier containing nucleic acid fragment, and application
  • Variant Gsdmd-C nucleic acid fragment, carrier containing nucleic acid fragment, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Construction and separation and purification of recombinant adeno-associated virus vector

[0059] gene sequence

[0060] Shown in SEQ ID NO:1 is the Gsdmd-C sequence,

[0061] Shown in SEQ ID NO: 2 is the amino acid sequence of Gsdmd-C,

[0062] Shown in SEQ ID NO:3 is the Left ITR sequence,

[0063] Shown in SEQ ID NO:4 is the CMV sequence,

[0064] Shown in SEQ ID NO:5 is the humanβ-globin intron sequence,

[0065] Shown in SEQ ID NO:6 is the MCS sequence,

[0066] Shown in SEQ ID NO:7 is hGHpA sequence,

[0067] Shown in SEQ ID NO:8 is the Right ITR sequence,

[0068] Shown in SEQ ID NO: 9 is the Ampicillin Resistance sequence,

[0069] Shown in SEQ ID NO:10 is the full nucleotide sequence,

[0070] Shown in SEQ ID NO:11 is f1 ori sequence,

[0071] Shown in SEQ ID NO: 12 is the recombinant sequence.

[0072] The construction of plasmid AAV_Gsdmd-C-HA, the characteristic map and the start and stop sites of each element are as follows: figure 1 A...

Embodiment 2

[0079] Example 2 Treatment of Retinal Detachment Model Mice with Recombinant Adeno-Associated Virus Vector

[0080] 1 Experimental materials

[0081] 1.1 Experimental animals

[0082] Normal Brown-Norway male rats were purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd., and the photoperiod was 12h light-12h dark. Animal Management Regulations.

[0083] 1.2 Experimental reagents and consumables

[0084] Normal saline (Zhejiang Tianrui Pharmaceutical Co., Ltd.), disposable injection needle (Becton Dickinson and Company, USA), HA antibody (Cell signaling technology), PI (Sigma Aldrich), secondary antibody goat anti-rabbit IgG (JacksonImmuno), goat Serum (Sigma), Polyethylene glycol octylphenyl ether (Triton X-100) (Sigma), Paraformaldehyde (Sigma).

[0085] 1.3 Experimental Instruments

[0086] Ophthalmology Experimental Operating Microscope (Nikon Company, Japan); Ophthalmic Microscopic Instruments (Suzhou Mingren Medical Instrument Co., Ltd.); Micro Sampler...

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Abstract

The invention relates to the field of biological medicine, particularly to a variant Gsdmd-C nucleic acid fragment, a carrier containing the nucleic acid fragment, and application. The nucleic acid fragment can realize better expression in a host (especially expression under coordination with an adeno-associated virus). In addition, the carrier containing the nucleic acid fragment has a good infection effect in rat photoreceptor cells, can realize stable expression in cell nucleuses, and can significantly reduce pyroptosis on a retina outer core layer and increase a survival rate of mouse photoreceptor cells, so that the fragment can be applied to preparing an agent which can treat pyroptosis caused by damage and apoptosis of photoreceptor cells in retinal detachment and has wide market prospect.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to a mutated Gsdmd-C nucleic acid fragment, the carrier and application of the nucleic acid fragment. Background technique [0002] Retinal detachment (RD) is a common pathological feature of many retinal choroidal diseases such as rhegmatogenous retinal detachment, age-related macular degeneration, and diabetic retinopathy. Retinal detachment is the separation of the retinal neuroepithelium from the retinal pigment epithelium (RPE) and choroid complex. Clinically, it is mainly manifested as vision loss, visual obstruction, floating objects in front of the eyes, and flashing sensation. When expanding to the macular area, the patient's visual acuity deteriorates rapidly. Since RPE and choroidal complexes are the main source of oxygen, blood, and energy for the outer retinal nerve fiber layer / photoreceptor cells, and the oxidative metabolism in the photoreceptor cells is active, in...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12N15/864A61K48/00A61K38/17A61P27/02
CPCA61K38/1709A61K48/005A61P27/02C07K14/47C12N15/86C12N2750/14143C12N2800/22
Inventor 汪枫桦
Owner 上海朗昇生物科技有限公司
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