智鼠故事 
B6-hRHO-P23H小鼠
复苏/繁育服务
产品名称
B6-hRHO-P23H
产品编号
C001495
品系全称
C57BL/6JCya-Rhotm2(hRHO*P23H)/Cya
品系背景
C57BL/6JCya
使用本品系发表的文献需注明： B6-hRHO-P23H mice (Catalog C001495) were purchased from Cyagen.
产品类型
周龄
性别
基因型
数量
品系介绍
视网膜色素变性（Retinitis Pigmentosa, RP）是一种遗传性视网膜疾病，全球患病率约为1:5000-1:3000。RP具有很大的临床和遗传异质性，视紫红质基因（Rhodopsin, RHO）突变导致约25％的显性RP [1]。RHO编码的视紫红蛋白与视觉光传导和GPCR下游信号密切相关，视紫红蛋白在视觉形成过程中光信号的传导至关重要，大多数RHO突变导致视紫红蛋白在感光细胞中高水平表达，使得大量的突变蛋白在细胞中定位异常并聚集，造成感光细胞凋亡，不能行使正常的光信号传导功能。此外，RHO基因的突变也与先天性静止性夜盲（CSNB）有关 [2-6]。目前靶向RHO基因以治疗视网膜色素变性的基因疗法有ASO、CRISPR等，全人源化动物模型的应用有助于推动RHO相关潜在疗法向临床试验进一步转化 [7-12]。
本模型为小鼠Rho基因人源化模型，利用基因编辑技术将小鼠内源性Rho基因替换为携带P23H突变的人源RHO基因片段，以在小鼠体内表达人源的视紫红蛋白。在该模型中，小鼠Rho基因被携带致病突变（P23H）的人源RHO基因取代，人源基因编码的异常蛋白在小鼠体内表达，因此该模型的视网膜外形和功能存在异常和视觉缺陷。此外，基于自主研发的TurboKnockout融合BAC重组的技术创新，赛业生物还可基于B6-hRHO全基因组人源化小鼠针对不同点突变提供定制服务，以满足广大研发人员关于视网膜色素变性（RP）疾病的药效学等实验需求。RHO基因突变可导致RHO介导的常染色体显性视网膜色素变性（RHO-adRP），在25%的常染色体显性遗传RP（adRP）病例中，有超过150种不同的RHO基因突变体。其中，P23H突变是常染色体显性遗传性视网膜色素变性的最常见原因之一，占adRP病例的10% [2]。之前的研究显示，携带该突变的杂合小鼠表现为与患者的疾病进展相似的视网膜病变和进行性视网膜变性 [3]，可用于视觉信号传导和视网膜色素变性（RP）的研究。B6-hRHO-P23H纯合小鼠发病比杂合小鼠更早且表型更严重。考虑到纯合小鼠后期失明导致的生长情况和生命周期的不确定性，建议使用B6-hRHO-P23H杂合小鼠进行实验。但也可按照特定的实验需求，选择纯合小鼠进行研究。
参考文献
Hartong, D. T., Berson, E. L., & Dryja, T. P. (2006). Retinitis pigmentosa. The Lancet, 368(9549), 1795-1809.
Meng D, Ragi SD, Tsang SH. Therapy in Rhodopsin-Mediated Autosomal Dominant Retinitis Pigmentosa. Mol Ther. 2020 Oct 7;28(10):2139-2149.
Sakami S, Maeda T, Bereta G, Okano K, Golczak M, Sumaroka A, Roman AJ, Cideciyan AV, Jacobson SG, Palczewski K. Probing mechanisms of photoreceptor degeneration in a new mouse model of the common form of autosomal dominant retinitis pigmentosa due to P23H opsin mutations. J Biol Chem. 2011 Mar 25;286(12):10551-67.
Dryja, T. P., McGee, T. L., Reichel, E., Hahn, L. B., Cowley, G. S., Yandell, D. W., ... & Berson, E. L. (1990). A point mutation of the rhodopsin gene in one form of retinitis pigmentosa. Nature, 343(6256), 364-366.
Zhang, X., Fu, W., Pang, C. P., & Yeung, K. Y. (2002). Screening for point mutations in rhodopsin gene among one hundred Chinese patients with retinitis pigmentosa. Zhonghua yi xue yi Chuan xue za zhi= Zhonghua Yixue Yichuanxue Zazhi= Chinese Journal of Medical Genetics, 19(6), 463-466.
Gamundi, M. J., Hernan, I., Muntanyola, M., Maseras, M., López‐Romero, P., Alvarez, R., ... & Carballo, M. (2008). Transcriptional expression of cis‐acting and trans‐acting splicing mutations cause autosomal dominant retinitis pigmentosa. Human mutation, 29(6), 869-878.
Biasutto, P., Adamson, P. S., Dulla, K., Murray, S., Monia, B., & McCaleb, M. (2019). Allele specific knock-down of human P23H rhodopsin mRNA and prevention of retinal degeneration in humanized P23H rhodopsin knock-in mouse, following treatment with an intravitreal GAPmer antisense oligonucleotide (QR-1123). Investigative Ophthalmology & Visual Science, 60(9), 5719-5719.
Editas Medicine, Inc. (2022, October 13). Press Release: Editas Medicine Presents Preclinical Data On EDIT-103 For Rhodopsin-Associated Autosomal Dominant Retinitis Pigmentosa At The European Society Of Gene And Cell Therapy Annual Meeting. Editasmedicine. https://ir.editasmedicine.com/news-releases/news-release-details/editas-medicine-presents-preclinical-data-edit-103-rhodopsin-0.
Patrizi, C., Llado, M., Benati, D., Iodice, C., Marrocco, E., Guarascio, R., ... & Recchia, A. (2021). Allele-specific editing ameliorates dominant retinitis pigmentosa in a transgenic mouse model. The American Journal of Human Genetics, 108(2), 295-308.
Li, P., Kleinstiver, B. P., Leon, M. Y., Prew, M. S., Navarro-Gomez, D., Greenwald, S. H., ... & Liu, Q. (2018). Allele-specific CRISPR-Cas9 genome editing of the single-base P23H mutation for rhodopsin-associated dominant retinitis pigmentosa. The CRISPR journal, 1(1), 55-64.
Liu, X., Jia, R., Meng, X., Li, Y., & Yang, L. (2022). Retinal degeneration in humanized mice expressing mutant rhodopsin under the control of the endogenous murine promoter. Experimental Eye Research, 215, 108893.
Wu, W. H., Tsai, Y. T., Huang, I. W., Cheng, C. H., Hsu, C. W., Cui, X., ... & Tsang, S. H. (2022). CRISPR genome surgery in a novel humanized model for autosomal dominant retinitis pigmentosa. Molecular Therapy, 30(4), 1407-1420.
ProQR Therapeutics. (2024). ProQR Receives Fast Track Designation from FDA for QR-1123 for Autosomal Dominant Retinitis Pigmentosa. Retrieved from ProQR Receives Fast Track Designation from FDA for QR-1123 for Autosomal Dominant https://www.proqr.com/press-releases/proqr-receives-fast-track-designation-from-fda-for-qr-1123-for-autosomal-dominant
构建方案
图1. B6-hRHO-P23H小鼠基因编辑打靶示意图。将小鼠的Rho基因ATG起始密码子至3'UTR下游部分片段替换为人源RHO基因ATG起始密码子至3'UTR下游部分片段，并将点突变p.P23H（CCC到CAC）引入人类RHO基因的1号外显子中。
