血管内皮生长因子受体 2 的基因编辑抑制体外血管生成

IF 3.7 4区 生物学 Q2 GENETICS & HEREDITY CRISPR Journal Pub Date : 2024-08-01 Epub Date: 2024-08-07 DOI:10.1089/crispr.2024.0019
Gaoen Ma, Hui Qi, Hongwei Deng, Lijun Dong, Qing Zhang, Junkai Ma, Yanhui Yang, Xiaohe Yan, Yajian Duan, Hetian Lei
{"title":"血管内皮生长因子受体 2 的基因编辑抑制体外血管生成","authors":"Gaoen Ma, Hui Qi, Hongwei Deng, Lijun Dong, Qing Zhang, Junkai Ma, Yanhui Yang, Xiaohe Yan, Yajian Duan, Hetian Lei","doi":"10.1089/crispr.2024.0019","DOIUrl":null,"url":null,"abstract":"<p><p>Vascular endothelial growth factor receptor (VEGFR)-2 is a key switch for angiogenesis, which is observed in various human diseases. In this study, a novel system for advanced prime editing (PE), termed PE6h, is developed, consisting of dual lentiviral vectors: (1) a clustered regularly interspaced palindromic repeat-associated protein 9 (H840A) nickase fused with reverse transcriptase and an enhanced PE guide RNA and (2) a dominant negative (DN) <i>MutL</i> homolog 1 gene with nicking guide RNA. PE6h was used to edit <i>VEGFR2</i> (c.18315T>A, 50.8%) to generate a premature stop codon (TAG from AAG), resulting in the production of DN-VEGFR2 (787 aa) in human retinal microvascular endothelial cells (HRECs). DN-VEGFR2 impeded VEGF-induced phosphorylation of VEGFR2, Akt, and extracellular signal-regulated kinase-1/2 and tube formation in PE6h-edited HRECs <i>in vitro</i>. Overall, our results highlight the potential of PE6h to inhibit angiogenesis <i>in vivo</i>.</p>","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":" ","pages":"188-196"},"PeriodicalIF":3.7000,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Prime Editing of Vascular Endothelial Growth Factor Receptor 2 Attenuates Angiogenesis <i>In Vitro</i>.\",\"authors\":\"Gaoen Ma, Hui Qi, Hongwei Deng, Lijun Dong, Qing Zhang, Junkai Ma, Yanhui Yang, Xiaohe Yan, Yajian Duan, Hetian Lei\",\"doi\":\"10.1089/crispr.2024.0019\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Vascular endothelial growth factor receptor (VEGFR)-2 is a key switch for angiogenesis, which is observed in various human diseases. In this study, a novel system for advanced prime editing (PE), termed PE6h, is developed, consisting of dual lentiviral vectors: (1) a clustered regularly interspaced palindromic repeat-associated protein 9 (H840A) nickase fused with reverse transcriptase and an enhanced PE guide RNA and (2) a dominant negative (DN) <i>MutL</i> homolog 1 gene with nicking guide RNA. PE6h was used to edit <i>VEGFR2</i> (c.18315T>A, 50.8%) to generate a premature stop codon (TAG from AAG), resulting in the production of DN-VEGFR2 (787 aa) in human retinal microvascular endothelial cells (HRECs). DN-VEGFR2 impeded VEGF-induced phosphorylation of VEGFR2, Akt, and extracellular signal-regulated kinase-1/2 and tube formation in PE6h-edited HRECs <i>in vitro</i>. Overall, our results highlight the potential of PE6h to inhibit angiogenesis <i>in vivo</i>.</p>\",\"PeriodicalId\":54232,\"journal\":{\"name\":\"CRISPR Journal\",\"volume\":\" \",\"pages\":\"188-196\"},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"CRISPR Journal\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1089/crispr.2024.0019\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/8/7 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"CRISPR Journal","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1089/crispr.2024.0019","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/7 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0

摘要

血管内皮生长因子受体(VEGFR)-2 是血管生成的关键开关,可在多种人类疾病中观察到。本研究开发了一种新型的高级质粒编辑(PE)系统,称为 PE6h,由双慢病毒载体组成:(1)融合了反转录酶和增强型 PE 引导 RNA 的簇状规则间隔 palindromic repeat-associated protein 9 (H840A)切口酶;(2)带有切口引导 RNA 的显性阴性 (DN) MutL 同源物 1 基因。利用 PE6h 编辑 VEGFR2(c.18315T>A,50.8%),生成一个过早终止密码子(从 AAG 变为 TAG),从而在人视网膜微血管内皮细胞(HRECs)中产生 DN-VEGFR2(787 aa)。DN-VEGFR2 阻碍了 VEGF 诱导的 VEGFR2、Akt 和细胞外信号调节激酶-1/2 的磷酸化,并阻碍了 PE6h 修饰的 HRECs 体外管形成。总之,我们的研究结果凸显了 PE6h 抑制体内血管生成的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Prime Editing of Vascular Endothelial Growth Factor Receptor 2 Attenuates Angiogenesis In Vitro.

Vascular endothelial growth factor receptor (VEGFR)-2 is a key switch for angiogenesis, which is observed in various human diseases. In this study, a novel system for advanced prime editing (PE), termed PE6h, is developed, consisting of dual lentiviral vectors: (1) a clustered regularly interspaced palindromic repeat-associated protein 9 (H840A) nickase fused with reverse transcriptase and an enhanced PE guide RNA and (2) a dominant negative (DN) MutL homolog 1 gene with nicking guide RNA. PE6h was used to edit VEGFR2 (c.18315T>A, 50.8%) to generate a premature stop codon (TAG from AAG), resulting in the production of DN-VEGFR2 (787 aa) in human retinal microvascular endothelial cells (HRECs). DN-VEGFR2 impeded VEGF-induced phosphorylation of VEGFR2, Akt, and extracellular signal-regulated kinase-1/2 and tube formation in PE6h-edited HRECs in vitro. Overall, our results highlight the potential of PE6h to inhibit angiogenesis in vivo.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
CRISPR Journal
CRISPR Journal Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.30
自引率
2.70%
发文量
76
期刊介绍: In recognition of this extraordinary scientific and technological era, Mary Ann Liebert, Inc., publishers recently announced the creation of The CRISPR Journal -- an international, multidisciplinary peer-reviewed journal publishing outstanding research on the myriad applications and underlying technology of CRISPR. Debuting in 2018, The CRISPR Journal will be published online and in print with flexible open access options, providing a high-profile venue for groundbreaking research, as well as lively and provocative commentary, analysis, and debate. The CRISPR Journal adds an exciting and dynamic component to the Mary Ann Liebert, Inc. portfolio, which includes GEN (Genetic Engineering & Biotechnology News) and more than 80 leading peer-reviewed journals.
期刊最新文献
Engineering CjCas9 for Efficient Base Editing and Prime Editing. CRISPR-Cas9-Mediated Targeting of Multidrug Resistance Genes in Methicillin-Resistant Staphylococcus aureus. Early Detection of Wildlife Disease Pathogens Using CRISPR-Cas System Methods. CRISPR-GRIT: Guide RNAs with Integrated Repair Templates Enable Precise Multiplexed Genome Editing in the Diploid Fungal Pathogen Candida albicans. Genome Editing in Apicomplexan Parasites: Current Status, Challenges, and Future Possibilities.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1