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一种实现绵羊FGF5基因精准突变的sgRNA、试剂盒和应用

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摘要

本发明提供了一种实现绵羊FGF5基因精准突变的sgRNA、试剂盒和应用，属于基因编辑技术领域，所述sgRNA由SEQ ID No.1所示的正向引物和SEQ ID No.2所示的反向引物退火获得；所述试剂盒，包括所述的sgRNA、FGF5‑单链寡核苷酸ssODN、Cas9 mRNA和DNA连接酶IV抑制剂SCR7。本发明提供的sgRNA、试剂盒及方法能够实现绵羊FGF5基因的高效、精准突变；能够用于绵羊育种，应用前景好。

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公开/公告号CN112553207A

专利类型发明专利
公开/公告日2021-03-26

原文格式PDF
申请/专利权人新疆畜牧科学院生物技术研究所(新疆畜牧科学院中国-澳大利亚绵羊育种研究中心);
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申请/专利号CN202011609515.3
发明设计人李文蓉;刘明军;刘晨曦;彭新荣;张雪梅;李忠慧;贺三刚;刘金瑞;张宁;玛依拉;
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申请日2020-12-30
分类号C12N15/113(20100101);C12N15/89(20060101);C12N15/10(20060101);C12N15/12(20060101);A01K67/027(20060101);
代理机构11385 北京方圆嘉禾知识产权代理有限公司;
代理人张秋菊
地址 830026 新疆维吾尔自治区乌鲁木齐市乌鲁木齐经济技术开发区阿里山街468号
入库时间 2023-06-19 10:25:58

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法律状态
2023-05-16

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1. 一种实现绵羊FGF5基因精准突变的sgRNA、试剂盒和应用 [P] . 中国专利： CN112553207A . 2021-03-26
2. 一种以RNA介导的特异性敲除FGF5基因获得基因编辑绵羊的方法及其专用sgRNA [P] . 中国专利： CN105132426B . 2018.01.09
3. method for specifically detecting the presence or absence of one or more single nucleotide mutations in one or more hiv-1 genes or proteins, method for detecting the presence or absence of mutations, selective oligonucleotide primer, oligonucleotide, kit for detecting a mutation , oligonucleotide blending for selective mutation detection and simultaneous detection of two or more mutations in one or more hiv-1 proteins [P] . BR112015029785A2 . 2017-09-26

机译：用于特异性地检测一种或多种hiv-1基因或蛋白质中一种或多种单核苷酸突变的存在或不存在的方法，用于检测突变的存在或不存在的方法，选择性寡核苷酸引物，寡核苷酸，用于检测突变的试剂盒，寡核苷酸掺合用于选择性突变检测并同时检测一种或多种hiv-1蛋白中的两个或多个突变
4. isolated polynucleotide, expression cassette, host cell, isolated polypeptide, non-human transgenic animal that has been altered to express an alkali resistant mutant polypeptide, an antibody that specifically binds to an alkali resistant mutant polypeptide, a mutation detection kit alk inhibitor resistance in a biological sample, and methods for evaluating a biological sample for an alk inhibitor resistance mutation to diagnose cancer that is resistant to or likely to develop resistance to at least one alkaline inhibitor. small alk molecule kinase, to evaluate a biological sample for an alk inhibitor resistance mutation, to diagnose cancer that is resistant to or likely to develop resistance to pf-02341066 in a patient, to specifically reduce expression of an alk-resistant mutant to treat cancer associated with activating aberrant loop capacity that is resistant to at least one small molecule alk kinase inhibitor treating an aberrant cancer that is resistant to pf-02341066, identifyin [P] . BR112012026239A2 . 2018-06-26

机译：分离的多核苷酸，表达盒，宿主细胞，分离的多肽，已被改变以表达耐碱突变体多肽的非人类转基因动物，与耐碱突变体多肽特异性结合的抗体，突变检测试剂盒生物样品，以及用于评估生物样品中是否存在对烷基抑制剂的抗性突变以诊断对至少一种碱性抑制剂有抗药性或可能发展出抗药性的癌症的方法。小分子激酶，用于评估生物学样品中是否具有抑制碱性磷酸酶抑制剂的突变，以诊断对患者具有抗药性或可能对pf-02341066产生抗药性的癌症，从而特异性降低抗癌突变体的表达以治疗癌症与激活抗至少一种小分子烷基激酶抑制剂的异常环路能力有关，该抑制剂治疗对pf-02341066有抵抗力的异常癌症
5. Polinucleotu00eddeo insulated tape of expression, host cell, isolated transgenic non-human animal polypeptide, which has been modified to express a polypeptide mutant with resistance to ALK, antibody that binds specifically to a polypeptide mutant with resistance to ALK, kit p For detection of a mutation with resistance to ALK inhibitor in a biological sample, and,Methods for evaluating a biological sample as a mutation with resistance to ALK inhibitor, for diagnosing a cancer that is resistant to or is likely to develop resistance to at least one inhibitor of kinase small molecule ALK, to evaluate a biological sample that Anto a mutation with resistance to ALK inhibitor.For diagnosing a cancer that is resistant to or is likely to develop resistance to at least one inhibitor of kinase small molecule ALK, to assess the biological sample as a mutation with resistance to ALK inhibitor, a cancer that is resistant to dianosticar Or is likely to develop resistance to PF - 02341066 in a pati [P] . BR112012026239A2 . 2016-11-29

机译：修饰的多聚核苷酸绝缘带，宿主细胞，分离的转基因非人类动物多肽，经过修饰以表达对ALK具有抗性的多肽突变体，与对ALK具有抗性的多肽突变体特异性结合的抗体，试剂盒p用于检测生物样品中对ALK抑制剂具有抗性的突变，以及用于评估生物样品为对ALK抑制剂具有抗性的突变的方法，用于诊断对至少一种对ALK抑制剂具有抗性或可能发展出抗性的癌症激酶小分子ALK，以评估Anto对ALK抑制剂具有抗性的突变的生物样品。用于诊断对至少一种激酶小分子ALK抑制剂具有抗药性或可能产生抗性的癌症，以评估该生物样品作为对ALK抑制剂有抗性的突变，即对狄诺司他抗性或可能对PF产生抗药性的癌症-02341066