WO2024021429A1 - 一种靶向视网膜的基因递送系统及其应用 - Google Patents
一种靶向视网膜的基因递送系统及其应用 Download PDFInfo
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- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
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- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
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Definitions
- the invention belongs to the field of biotechnology, and specifically relates to a retina-targeted gene delivery system and its application.
- Retinal diseases seriously endanger human health, but there is a lack of treatment methods. Although some progress has been made using viral vector-based gene therapy in recent years, it only targets a small number of genetic diseases and needs longer clinical evaluation. It has been found that a variety of retinal diseases are related to bipolar cells, or are accompanied by bipolar cell lesions during their occurrence, leading to the aggravation of the disease, such as neuronal ceroid lipofuscinosis (Neuronal ceroid lipofuscinosis). lipofuscinosis (NCL), retinoschisis and retinitis pigmentosa (RP) caused by RS1 gene mutations, etc.
- NCL neuronal ceroid lipofuscinosis
- RP retinoschisis
- RP retinitis pigmentosa
- the present invention provides a retina-targeted gene delivery system and its application.
- the present invention provides a gene delivery system targeting the retina.
- the retina-targeted gene delivery system includes an adeno-associated virus vector rAAV11, which includes a gene encoding a marker protein and a promoter, and the adeno-associated virus vector rAAV11 targets retinal cells.
- the marker protein in the gene encoding the marker protein is selected from fluorescent proteins and/or enzyme reaction chromogenic proteins
- the fluorescent protein is selected from BFP, CFP, GFP, YFP, RFP, iRFP, Cerulean, Venus, eGFP , one or more of eCFP, eYFP, eBFP, DsRed, dTomato, tdTomato, mCherry, mKate, mApple, mBanana, mCitrine, mOrange, mPlum, tagRFP and tagBFP
- the enzyme reaction chromogenic protein is selected from HRP, Firefly One or more of luciferase and Renilla luciferase.
- the promoter is selected from CAG, CMV, hUbC, Ef1 ⁇ , nEF, hSyn, CaMKII ⁇ , Vgat, Thy1, TRE, UAS, GFAP, gfaABC1D, TH, RPE65, TRE, CBA, PGK, E-SARE, C -fos, RAM, SST, PV, mDlX, NPY, CR, TCAP, SFRP2, ChAT, TPH2, mTH, GAD67, GFAP104, CD68, Nestin, MBP, TRPV1, L7/Pcp2, mOXT, RK, hGRK1, CAR, Grm6 , one or more of ROH, Nrl, MCK, dMCK and tMCK.
- the retinal cells are selected from neurons and/or glial cells, the neurons are selected from one or both bipolar cells and photoreceptor cells, and the glial cells are selected from ganglion cells. .
- An object of the present invention is to provide a method for labeling retinal neural networks.
- S1 Inject the adeno-associated virus vector into the vitreous cavity of the eye
- the adeno-associated virus vector is rAAV11, and the adeno-associated virus vector includes a gene encoding a marker protein and a promoter.
- the marker protein in the gene encoding the marker protein is selected from fluorescent proteins and/or enzyme reaction chromogenic proteins
- the fluorescent protein is selected from BFP, CFP, GFP, YFP, RFP, iRFP, Cerulean, Venus, eGFP , one or more of eCFP, eYFP, eBFP, DsRed, dTomato, tdTomato, mCherry, mKate, mApple, mBanana, mCitrine, mOrange, mPlum, tagRFP and tagBFP
- the enzyme reaction chromogenic protein is selected from HRP, Firefly One or more of luciferase and Renilla luciferase.
- the promoter is selected from CAG, CMV, hUbC, Ef1 ⁇ , nEF, hSyn, CaMKII ⁇ , Vgat, Thy1, TRE, UAS, GFAP, gfaABC1D, TH, RPE65, TRE, CBA, PGK, E-SARE, C -fos, RAM, SST, PV, mDlX, NPY, CR, TCAP, SFRP2, ChAT, TPH2, mTH, GAD67, GFAP104, CD68, Nestin, MBP, TRPV1, L7/Pcp2, mOXT, RK, hGRK1, CAR, Grm6 , one or more of ROH, Nrl, MCK, dMCK and tMCK.
- An object of the present invention is to provide an application of an adeno-associated virus vector to express a target protein or functional RNA in retinal neurons, or to prepare a reagent for expressing a target protein or functional RNA in retinal neurons.
- the adeno-associated virus vector in expressing a protein of interest or functional RNA in retinal neurons, or use in preparing a reagent for expressing a protein of interest or functional RNA in retinal neurons, wherein the adeno-associated virus vector It is rAAV11, and the adeno-associated virus vector includes genes and promoters encoding target proteins or functional RNAs.
- the target proteins are selected from marker proteins and/or active proteins.
- the active proteins are selected from the group consisting of activating neuronal proteins and inhibitory proteins.
- the functional RNA is selected from small RNA, small interfering RNA, small hairpin RNA, small guide RNA, organelle localization RNA and RNA sequencing or in situ hybridization analysis.
- small RNA small interfering RNA
- small hairpin RNA small guide RNA
- organelle localization RNA RNA sequencing or in situ hybridization analysis.
- Barcode RNA One or more of Barcode RNA.
- One object of the present invention is to provide a method for expressing target proteins or functional RNA in retinal neurons.
- the method of expressing a target protein or functional RNA in retinal neurons includes injecting an adeno-associated virus vector into the vitreous cavity of the eye.
- the adeno-associated virus vector is rAAV11, and the adeno-associated virus vector also contains a protein encoding the target protein. or functional RNA genes and promoters,
- the target protein is selected from marker proteins and/or active proteins
- the active protein is selected from neuron-activating proteins, neuron-inhibiting proteins, calcium ion signal probe proteins, small molecule signal probe proteins, and apoptosis-mediating proteins.
- proteins, disease-related mutant proteins, normal proteins under physiological conditions, cytokines, antiviral factors, viral infection co-receptors, recombinases and gene editing tool proteins and the functional RNA is selected from small One or more of RNA, small interfering RNA, small hairpin RNA, small guide RNA, organelle-localized RNA, and Barcode RNA for RNA sequencing or in situ hybridization analysis.
- One object of the present invention is to provide an application of an adeno-associated virus vector in the preparation of drugs for treating diseases caused by retinal neuron abnormalities.
- adeno-associated virus vector in the preparation of drugs for treating diseases caused by retinal neuron abnormalities, wherein the adeno-associated virus vector is rAAV11, and the adeno-associated virus vector also contains a gene encoding a therapeutic protein or functional RNA genes and promoters,
- the target protein is selected from marker proteins and/or active proteins
- the active protein is selected from activating neuron proteins, inhibiting neuron proteins, normal proteins under physiological conditions, cytokines, antiviral factors and gene editing tool proteins.
- the functional RNA is selected from one or more of small RNA, small interfering RNA, small hairpin RNA and small guide RNA.
- the present invention has discovered a new use of the adeno-associated virus rAAV11 vector.
- the rAAV11 vector can express the target gene in the neurons of the retina. Based on this, the rAAV11 vector can carry the target gene for labeling and controlling the retinal nervous system, and can also be used as a gene therapy. carrier.
- rAAV11 vector can deliver genes to retinal bipolar cells efficiently and specifically.
- Figure 1 shows the labeling results of rAAV11-Ef1 ⁇ -eGFP injected into the vitreous cavity.
- the pictures marked 1 and 2 in Figure 1 represent infected bipolar cells.
- a method of delivering genes to the retina has been developed using the adeno-associated virus rAAV11 vector.
- Adeno-associated virus rAAV11 vector can label retinal nerve cells.
- rAAV11-Ef1 ⁇ -eGFP was injected into the vitreous cavity of the eyes of C57BL/6J mice, and neuron cell bodies labeled with green fluorescent protein were observed in the retina. The results showed that rAAV11 can pass through the glass. Intracorporeal injection of retinal-infected nerve cells.
- the injection method involved in the present invention can be subretinal injection or suprachoroidal injection.
- the adeno-associated virus vector rAAV11 is a recombinant virus obtained through three-plasmid packaging. See AAV11 permits efficient retrograde targeting of projection neurons, Han et al. bioRxiv 2022.01.13.476170; doi: https://doi.org/10.1101/2022.01.13.476170.
- AAV11 permits efficient retrograde targeting of projection neurons, Han et al. bioRxiv 2022.01.13.476170; doi: https://doi.org/10.1101/2022.01.13.476170, using a three-plasmid packaging system to package adeno-associated virus, rAAV11 vector is packaged for virus, and iodixanol gradient centrifugation is used for concentration and purification, and finally SYBR Green qPCR is used The titer of the recombinant adeno-associated virus was detected using the method, and the obtained titer was 8.7 ⁇ 1012 vg/mL.
- C57BL/6J mice about 2 months old purchased from Hunan Slack Jingda Experimental Animal Co., Ltd.
- the cells were killed by neck dissection and the eyeballs were quickly removed.
- the tissue was embedded in embedding agent, cut into 20 ⁇ m thick slices with a freezing microtome, and mounted while cutting. After staining with DAPI, the slices were imaged using a slide scanner.
- rAAV11-Ef1 ⁇ -eGFP can infect retinal photoreceptor cells, bipolar cells and ganglion cells, but mainly infects bipolar cells.
- Pictures labeled 1 and 2 in the figure represent infected bipolar cells.
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Abstract
本发明属于生物技术领域，具体涉及靶向视网膜的基因递送系统及其应用。该靶向视网膜的基因递送系统包括腺相关病毒载体rAAV11，所述腺相关病毒载体rAAV11包括编码标记蛋白的基因和启动子，所述腺相关病毒载体rAAV11靶向视网膜细胞。相比于其他载体，rAAV11载体可以高效且特异地将基因递送到视网膜双极细胞。
Description
本发明属于生物技术领域，具体涉及靶向视网膜的基因递送系统及其应用。
视网膜疾病严重危害人类健康，但缺乏治疗手段。虽然近几年利用基于病毒载体的基因治疗取得一些进展，但仅针对少部分遗传性疾病，且有待更长时间的临床评估。现已发现多种视网膜疾病与双极细胞有关，或者在其发生过程伴随双极细胞的病变而导致疾病的加重，例如神经元蜡样脂褐质沉积症(Neuronal ceroid
lipofuscinosis，NCL)、RS1基因突变的视网膜劈裂症和视网膜色素变性(Retinitis pigmentosa ,RP)等。然而，双极细胞的病变在疾病中的作用机制尚不清楚，严重阻碍了这类视网膜疾病的临床诊断和基因治疗，其中原因包括高效特异靶向这类细胞的研究工具的缺乏，因此有必要开发高效靶向视网膜，特别是视网膜双极细胞的工具病毒载体。
鉴于此，本发明提供了一种靶向视网膜的基因递送系统及其应用。
为了解决高效特异靶向这类细胞的研究工具的缺乏的问题，本发明提供了一种靶向视网膜的基因递送系统。
该靶向视网膜的基因递送系统包括腺相关病毒载体rAAV11，所述腺相关病毒载体rAAV11包括编码标记蛋白的基因和启动子，所述腺相关病毒载体rAAV11靶向视网膜细胞。
进一步地，所述编码标记蛋白的基因中的标记蛋白选自荧光蛋白和/或酶反应显色蛋白，所述荧光蛋白选自BFP、CFP、GFP、YFP、RFP、iRFP、Cerulean、Venus、eGFP、eCFP、eYFP、eBFP、DsRed、dTomato、tdTomato、mCherry、mKate、mApple、mBanana、mCitrine、mOrange、mPlum、tagRFP和tagBFP中的一种或多种，所述酶反应显色蛋白选自HRP、Firefly
luciferase和Renilla luciferase中的一种或多种。
进一步地，所述启动子选自CAG、CMV、hUbC、Ef1α、nEF、hSyn、CaMKIIα、Vgat、Thy1、TRE、UAS、GFAP、gfaABC1D、TH、RPE65、TRE、CBA、PGK、E-SARE、C-fos、RAM、SST、PV、mDlX、NPY、CR、TCAP、SFRP2、ChAT、TPH2、mTH、GAD67、GFAP104、CD68、Nestin、MBP、TRPV1、L7/Pcp2、mOXT、RK、hGRK1、CAR、Grm6、ROH、Nrl、MCK、dMCK和tMCK中的一种或多种。
进一步地，所述视网膜细胞选自神经元和/或神经胶质细胞，所述神经元选自双极细胞和感光细胞中的一种或两种，所述神经胶质细胞选自神经节细胞。
本发明的一个目的是提供一种视网膜神经网络的标记方法。
该视网膜神经网络的标记方法包括以下步骤：
S1：将腺相关病毒载体注射至眼睛的玻璃体腔中；
S2：观察视网膜中标记蛋白的分布情况；
其中，所述腺相关病毒载体为rAAV11，所述关病毒载体包括编码标记蛋白的基因和启动子。
进一步地，所述编码标记蛋白的基因中的标记蛋白选自荧光蛋白和/或酶反应显色蛋白，所述荧光蛋白选自BFP、CFP、GFP、YFP、RFP、iRFP、Cerulean、Venus、eGFP、eCFP、eYFP、eBFP、DsRed、dTomato、tdTomato、mCherry、mKate、mApple、mBanana、mCitrine、mOrange、mPlum、tagRFP和tagBFP中的一种或多种，所述酶反应显色蛋白选自HRP、Firefly
luciferase和Renilla luciferase中的一种或多种。
进一步地，所述启动子选自CAG、CMV、hUbC、Ef1α、nEF、hSyn、CaMKIIα、Vgat、Thy1、TRE、UAS、GFAP、gfaABC1D、TH、RPE65、TRE、CBA、PGK、E-SARE、C-fos、RAM、SST、PV、mDlX、NPY、CR、TCAP、SFRP2、ChAT、TPH2、mTH、GAD67、GFAP104、CD68、Nestin、MBP、TRPV1、L7/Pcp2、mOXT、RK、hGRK1、CAR、Grm6、ROH、Nrl、MCK、dMCK和tMCK中的一种或多种。
本发明的一个目的是提供一种腺相关病毒载体的在视网膜神经元中表达目的蛋白或功能性RNA的应用，或者在制备在视网膜神经元中表达目的蛋白或功能性RNA的试剂中的应用。
该腺相关病毒载体的在视网膜神经元中表达目的蛋白或功能性RNA的应用，或者在制备在视网膜神经元中表达目的蛋白或功能性RNA的试剂中的应用，其中，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中包括编码目的蛋白或功能性RNA的基因和启动子，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、钙离子信号探针蛋白、小分子信号探针蛋白、介导凋亡蛋白、疾病相关突变蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子、病毒感染辅助受体、重组酶和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA、小向导RNA、细胞器定位RNA和用于RNA测序或原位杂交分析的Barcode RNA中的一种或多种。
本发明的一个目的是提供一种在视网膜神经元中表达目的蛋白或功能性RNA的方法。
该在视网膜神经元中表达目的蛋白或功能性RNA的方法包括将腺相关病毒载体注射至眼睛的玻璃体腔中，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中还包含编码目的蛋白或功能性RNA的基因和启动子，
其中，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、钙离子信号探针蛋白、小分子信号探针蛋白、介导凋亡蛋白、疾病相关突变蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子、病毒感染辅助受体、重组酶和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA、小向导RNA、细胞器定位RNA和用于RNA测序或原位杂交分析的Barcode RNA中的一种或多种。
本发明的一个目的是提供一种腺相关病毒载体在制备治疗视网膜神经元异常导致的疾病的药物中的应用。
该腺相关病毒载体在制备治疗视网膜神经元异常导致的疾病的药物中的应用，其中，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中还包含编码治疗用蛋白或功能性RNA的基因和启动子，
其中，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA和小向导RNA中的一种或多种
本发明发现了腺相关病毒rAAV11载体的新用途，rAAV11载体能够将目的基因表达在视网膜的神经元中，基于此rAAV11载体可以携带目的基因用于标记、操控视网膜神经系统，也可以作为基因治疗的载体。
相比于其他载体，rAAV11载体可以高效且特异地将基因递送到视网膜双极细胞。
图1为将rAAV11-Ef1α-eGFP注射到玻璃体腔的标记结果，图1中标记为1和2的图片代表感染的双极细胞。
为了使本发明的上述目的、特征和优点能够更加明显易懂，下面结合附图对本发明的具体实施方式做详细的说明，但不能理解为对本发明的可实施范围的限定。
在本发明中，提供了利用腺相关病毒rAAV11载体开发了一种递送基因到视网膜的方法。
腺相关病毒rAAV11载体能够标记视网膜的神经细胞。在本发明的具体实施案例中，将rAAV11-Ef1α-eGFP注射至C57BL/6J品系小鼠眼睛的玻璃体腔中，在视网膜中观察到被绿色荧光蛋白标记的神经元胞体，结果表明rAAV11可以通过玻璃体腔注射感染视网膜的神经细胞。本发明中涉及的注射方法可以为视网膜下注射或脉络膜上注射。
在本发明中，腺相关病毒载体rAAV11为通过三质粒包装获得的重组病毒。参见AAV11 permits efficient
retrograde targeting of projection neurons，Han et al. bioRxiv
2022.01.13.476170; doi: https://doi.org/10.1101/2022.01.13.476170。
实施例1重组腺相关病毒的制备
采用已知方法获得重组腺相关病毒rAAV11，具体参见
AAV11 permits efficient retrograde targeting of projection neurons，Han
et al. bioRxiv 2022.01.13.476170; doi:
https://doi.org/10.1101/2022.01.13.476170，采用三质粒包装系统包装腺相关病毒方法，对rAAV11载体进行病毒包装，并用碘克沙醇梯度离心法进行浓缩和纯化，最后用SYBR Green qPCR法检测重组腺相关病毒滴度，获得滴度为8.7×1012 vg/mL。
实施例2重组腺相关病毒标记视网膜神经细胞的活体测试
2个月龄左右的C57BL/6J小鼠（购自湖南斯莱克景达实验动物有限公司）通过腹腔注射1%戊巴比妥钠麻醉后，在其眼睛的玻璃体腔位置注射1.5
µL重组腺相关病毒rAAV11-Ef1α-eGFP，病毒经过3周的表达后，断颈处死，并快速取眼球。放入4%的PFA溶液并置于冰上固定20min后，在角膜上剪口，然后放入4度冰箱固定2小时后，再用30%的蔗糖-PBS溶液脱水，将脱水后的组织用组织包埋剂包埋，用冰冻切片机切成20 μm厚的片子，边切边贴片，经DAPI染色后使用玻片扫描仪对其进行成像。
活体检测结果如图1所示，rAAV11-Ef1α-eGFP可以感染视网膜的感光细胞、双极细胞和神经节细胞，但主要感染双极细胞。图中标记为1和2的图片代表感染的双极细胞。
Claims (10)
- 一种靶向视网膜的基因递送系统，其特征在于，包括腺相关病毒载体rAAV11，所述腺相关病毒载体rAAV11包括编码标记蛋白的基因和启动子，所述腺相关病毒载体rAAV11靶向视网膜细胞。
- 如权利要求1所述的靶向视网膜的基因递送系统，其特征在于，所述编码标记蛋白的基因中的标记蛋白选自荧光蛋白和/或酶反应显色蛋白，所述荧光蛋白选自BFP、CFP、GFP、YFP、RFP、iRFP、Cerulean、Venus、eGFP、eCFP、eYFP、eBFP、DsRed、dTomato、tdTomato、mCherry、mKate、mApple、mBanana、mCitrine、mOrange、mPlum、tagRFP和tagBFP中的一种或多种，所述酶反应显色蛋白选自HRP、Firefly luciferase和Renilla luciferase中的一种或多种。
- 如权利要求1所述的靶向视网膜的基因递送系统，其特征在于，所述启动子选自CAG、CMV、hUbC、Ef1α、nEF、hSyn、CaMKIIα、Vgat、Thy1、TRE、UAS、GFAP、gfaABC1D、TH、RPE65、TRE、CBA、PGK、E-SARE、C-fos、RAM、SST、PV、mDlX、NPY、CR、TCAP、SFRP2、ChAT、TPH2、mTH、GAD67、GFAP104、CD68、Nestin、MBP、TRPV1、L7/Pcp2、mOXT、RK、hGRK1、CAR、Grm6、ROH、Nrl、MCK、dMCK和tMCK中的一种或多种。
- 如权利要求1所述的靶向视网膜的基因递送系统，其特征在于，所述视网膜细胞选自神经元和/或神经胶质细胞，所述神经元选自双极细胞和感光细胞中的一种或两种，所述神经胶质细胞选自神经节细胞。
- 一种视网膜神经网络的标记方法，其特征在于，包括以下步骤：S1：将腺相关病毒载体注射至眼睛的玻璃体腔中；S2：观察视网膜中标记蛋白的分布情况；其中，所述腺相关病毒载体为rAAV11，所述关病毒载体包括编码标记蛋白的基因和启动子。
- 如权利要求5所述的视网膜神经网络的标记方法，其特征在于，所述编码标记蛋白的基因中的标记蛋白选自荧光蛋白和/或酶反应显色蛋白，所述荧光蛋白选自BFP、CFP、GFP、YFP、RFP、iRFP、Cerulean、Venus、eGFP、eCFP、eYFP、eBFP、DsRed、dTomato、tdTomato、mCherry、mKate、mApple、mBanana、mCitrine、mOrange、mPlum、tagRFP和tagBFP中的一种或多种，所述酶反应显色蛋白选自HRP、Firefly luciferase和Renilla luciferase中的一种或多种。
- 如权利要求5所述的视网膜神经网络的标记方法，其特征在于，所述启动子选自CAG、CMV、hUbC、Ef1α、nEF、hSyn、CaMKIIα、Vgat、Thy1、TRE、UAS、GFAP、gfaABC1D、TH、RPE65、TRE、CBA、PGK、E-SARE、C-fos、RAM、SST、PV、mDlX、NPY、CR、TCAP、SFRP2、ChAT、TPH2、mTH、GAD67、GFAP104、CD68、Nestin、MBP、TRPV1、L7/Pcp2、mOXT、RK、hGRK1、CAR、Grm6、ROH、Nrl、MCK、dMCK和tMCK中的一种或多种。
- 腺相关病毒载体的在视网膜神经元中表达目的蛋白或功能性RNA的应用，或者在制备在视网膜神经元中表达目的蛋白或功能性RNA的试剂中的应用，其特征在于，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中包括编码目的蛋白或功能性RNA的基因和启动子，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、钙离子信号探针蛋白、小分子信号探针蛋白、介导凋亡蛋白、疾病相关突变蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子、病毒感染辅助受体、重组酶和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA、小向导RNA、细胞器定位RNA和用于RNA测序或原位杂交分析的Barcode RNA中的一种或多种。
- 在视网膜神经元中表达目的蛋白或功能性RNA的方法，其特征在于，包括将腺相关病毒载体注射至眼睛的玻璃体腔中，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中还包含编码目的蛋白或功能性RNA的基因和启动子，其中，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、钙离子信号探针蛋白、小分子信号探针蛋白、介导凋亡蛋白、疾病相关突变蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子、病毒感染辅助受体、重组酶和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA、小向导RNA、细胞器定位RNA和用于RNA测序或原位杂交分析的Barcode RNA中的一种或多种。
- 腺相关病毒载体在制备治疗视网膜神经元异常导致的疾病的药物中的应用，其特征在于，所述腺相关病毒载体为rAAV11，所述腺相关病毒载体中还包含编码治疗用蛋白或功能性RNA的基因和启动子，其中，所述目的蛋白选自标记蛋白和/或活性蛋白，所述活性蛋白选自激活神经元蛋白、抑制神经元蛋白、生理条件下的正常蛋白、细胞因子、抗病毒因子和基因编辑工具蛋白中的一种或多种，所述功能性RNA选自小RNA、小干扰RNA、小发卡RNA和小向导RNA中的一种或多种。
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- 2022-12-07 WO PCT/CN2022/137380 patent/WO2024021429A1/zh unknown
Patent Citations (4)
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|---|---|---|---|---|
| US20120093772A1 (en) * | 2008-05-20 | 2012-04-19 | Alan Horsager | Vectors for delivery of light sensitive proteins and methods of use |
| US20220226507A1 (en) * | 2019-05-17 | 2022-07-21 | Research Institute At Nationwide Children's Hospital | Optimized gene therapy targeting retinal cells |
| CN114728132A (zh) * | 2019-09-20 | 2022-07-08 | 梅拉格特克斯治疗学股份有限公司 | 注射系统及其使用方法 |
| CN115323001A (zh) * | 2022-07-28 | 2022-11-11 | 深圳先进技术研究院 | 一种靶向视网膜的基因递送系统及其应用 |
Non-Patent Citations (1)
| Title |
|---|
| HAN ZENGPENG, LUO NENGSONG, KOU JIAXIN, LI LEI, MA WENYU, PENG SIQI, XU ZIHONG, ZHANG WEN, QIU YUXIANG, WU YANG, WANG JIE, YE CHAO: "AAV11 permits efficient retrograde targeting of projection neurons", BIORXIV, 13 January 2022 (2022-01-13), XP093134147, DOI: 10.1101/2022.01.13.476170 * |
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