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精准调控CRISPR/Cas9基因编辑技术研究进展 被引量:13
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作者 曹俊霞 王友亮 王征旭 《遗传》 CAS CSCD 北大核心 2020年第12期1168-1177,共10页
基因编辑(gene editing)是一种能对细胞和生物体基因组一小段DNA进行定点修饰或者删除、插入的基因工程技术。基因编辑技术在疾病治疗、基因功能调控、基因检测、药物研发和作物育种等方面具有广阔的应用前景,但在应用中也逐渐显现出脱... 基因编辑(gene editing)是一种能对细胞和生物体基因组一小段DNA进行定点修饰或者删除、插入的基因工程技术。基因编辑技术在疾病治疗、基因功能调控、基因检测、药物研发和作物育种等方面具有广阔的应用前景,但在应用中也逐渐显现出脱靶、基因毒性等副作用问题。CRISPR(clustered regularly interspaced short palindromic repeats)系统中核酸酶Cas9蛋白能通过与gRNA(guide RNA)结合特异性识别靶DNA并进行酶切反应,由于Cas9蛋白和gRNA在其自身活性、识别位点及结合能力等方面具有不同的特性,因此在应用中可以通过对Cas9蛋白酶的活性以及与靶DNA在时间和空间上的结合进行精准调控,主要调控方法包括使用光、温度和药物等调节Cas9融合蛋白、抗CRISPR蛋白、核酸类和小分子类化合物抑制剂的使用等,从而能有效地防范基因编辑技术的风险和增强精准调控基因编辑技术的实际应用性。本文就目前如何精准调控基因编辑技术,尤其是精准调控CRISPR/Cas9基因编辑技术的方法进行了综述,以期为人类疾病治疗、作物育种、家畜遗传改良和防范生物技术缪用等提供借鉴和研究思路。 展开更多
关键词 基因编辑 基因编辑调控 抗CRISPR蛋白 小分子抑制剂
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基因编辑技术在军事医学领域的潜在应用
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作者 王征旭 曹俊霞 王友亮 《解放军医学院学报》 CAS 北大核心 2023年第11期1187-1193,共7页
基因编辑技术是一种对生物体目标基因进行定点“编辑”的工程技术,近年来发展非常迅速,已冲击到生命科学的各个领域,是世界范围内竞争最为激烈的下一代核心颠覆性生物技术。基因编辑技术已应用于农作物改良、畜牧业育种、遗传与代谢性... 基因编辑技术是一种对生物体目标基因进行定点“编辑”的工程技术,近年来发展非常迅速,已冲击到生命科学的各个领域,是世界范围内竞争最为激烈的下一代核心颠覆性生物技术。基因编辑技术已应用于农作物改良、畜牧业育种、遗传与代谢性疾病的治疗、抗细菌和病毒感染、药物筛选、制备新型生物燃料等领域。其在军事医学领域也有重要用途:通过基因编辑技术,使病原体危害性增大、毒性增强;靶向病原体的保守序列设计药物、疫苗等,可以防御多个毒株和变异体的感染;靶向宿主,CRISPR/dCas9系统介导的表观遗传基因编辑,不改变DNA遗传性状,也能可逆性增强人体效能,抵抗生物、化学、放射损伤;靶向基因编辑器自身的核酸酶和引导RNA,能精准调控基因编辑技术,防御基因编辑技术谬用和新型基因类生物战剂造成的危害。基因编辑技术在传染病诊治领域也具有显著优势,能彻底治愈某些类型的传染病。 展开更多
关键词 基因编辑技术 基因增强 表观遗传基因编辑 基因编辑调控技术
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CRISPR/Cas9在放线菌合成生物学研究中的应用和发展 被引量:1
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作者 黄蓉 段燕文 朱湘成 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2019年第9期960-967,共8页
放线菌是活性天然产物和抗生素药物的重要来源。利用合成生物学高效地开发其中丰富的天然产物资源,将为加速新药开发奠定坚实的基础。CRISPR/Cas9作为一种多功能基因编辑系统,因其便捷高效而被广泛应用于真核生物的遗传操作。但在原核... 放线菌是活性天然产物和抗生素药物的重要来源。利用合成生物学高效地开发其中丰富的天然产物资源,将为加速新药开发奠定坚实的基础。CRISPR/Cas9作为一种多功能基因编辑系统,因其便捷高效而被广泛应用于真核生物的遗传操作。但在原核生物尤其是放线菌中的应用仍处于起步阶段,机遇和挑战并存。本综述总结了目前CRISPR/Cas9系统在放线菌基因编辑和调控,以及活性天然产物的产量提升、生物合成机制解析和资源开发等方面的研究进展。同时,也对该系统在应用中面临的包括重组修复效率低,以及靶向切割效率不足等关键挑战进行了分析,并提出了相应的优化解决方法。随着CRISPR/Cas9在放线菌应用中的不断完善和发展,将极大地推动放线菌的合成生物学研究,促进其中天然产物资源的有效挖掘和应用开发。 展开更多
关键词 放线菌 CRISPR/Cas9 合成生物学 基因编辑调控 微生物天然产物
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Gene editing: an instrument for practical application of gene biology to plant breeding 被引量:2
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作者 Yuan-yuan TAN Hao DU +5 位作者 Xia WU Yan-hua LIU Meng JIANG Shi-yong SONG Liang WU Qing-yao SHU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2020年第6期460-473,共14页
Plant breeding is well recognized as one of the most important means to meet food security challenges caused by the ever-increasing world population. During the past three decades, plant breeding has been empowered by... Plant breeding is well recognized as one of the most important means to meet food security challenges caused by the ever-increasing world population. During the past three decades, plant breeding has been empowered by both new knowledge on trait development and regulation(e.g., functional genomics) and new technologies(e.g., biotechnologies and phenomics). Gene editing, particularly by clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein(Cas) and its variants, has become a powerful technology in plant research and may become a game-changer in plant breeding. Traits are conferred by coding and non-coding genes. From this perspective, we propose different editing strategies for these two types of genes. The activity of an encoded enzyme and its quantity are regulated at transcriptional and post-transcriptional, as well as translational and post-translational, levels. Different strategies are proposed to intervene to generate gene functional variations and consequently phenotype changes. For non-coding genes, trait modification could be achieved by regulating transcription of their own or target genes via gene editing. Also included is a scheme of protoplast editing to make gene editing more applicable in plant breeding. In summary, this review provides breeders with a host of options to translate gene biology into practical breeding strategies, i.e., to use gene editing as a mechanism to commercialize gene biology in plant breeding. 展开更多
关键词 Gene editing Expression regulation Novel allele Trait development Plant breeding
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Juvenile hormone membrane signaling phosphorylates USP and thus potentiates 20-hydroxyecdysone action in Drosophila 被引量:4
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作者 Yue Gao Suning Liu +8 位作者 Qiangqiang Jia Lixian Wu Dongwei Yuan Emma Y.Li Qili Feng Guirong Wang Subba R.Palli Jian Wang Sheng Li 《Science Bulletin》 SCIE EI CSCD 2022年第2期186-197,M0004,共13页
Juvenile hormone(JH) and 20-hydroxyecdysone(20 E) coordinately regulate development and metamorphosis in insects. Two JH intracellular receptors, methoprene-tolerant(Met) and germ-cell expressed(Gce), have been identi... Juvenile hormone(JH) and 20-hydroxyecdysone(20 E) coordinately regulate development and metamorphosis in insects. Two JH intracellular receptors, methoprene-tolerant(Met) and germ-cell expressed(Gce), have been identified in the fruit fly Drosophila melanogaster. To investigate JH membrane signaling pathway without the interference from JH intracellular signaling, we characterized phosphoproteome profiles of the Met gce double mutant in the absence or presence of JH in both chronic and acute phases.Functioning through a potential receptor tyrosine kinase and phospholipase C pathway, JH membrane signaling activated protein kinase C(PKC) which phosphorylated ultraspiracle(USP) at Ser35, the PKC phosphorylation site required for the maximal action of 20 E through its nuclear receptor complex Ec RUSP. The usp;mutant, in which Ser was replaced with Ala at position 35 by genome editing, showed decreased expression of Halloween genes that are responsible for ecdysone biosynthesis and thus attenuated 20 E signaling that delayed developmental timing. The usp;mutant also showed lower Yorkie activity that reduced body size. Altogether, JH membrane signaling phosphorylates USP at Ser35 and thus potentiates 20 E action that regulates the normal fly development. This study helps better understand the complex JH signaling network. 展开更多
关键词 Juvenile hormone Receptor tyrosine kinase PHOSPHOPROTEOMICS USP 20-Hydroxyecdysone action Yorkie activity
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