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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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[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 detachmentphotoreceptor cell injury and death.
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Embodiment 1
[0058] Example 1 Construction and separation and purification of recombinant adeno-associated virus vector
[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-globinintron 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.
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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 retinaouter 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...
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