首页> 外国专利> IN VIVO RNA OR PROTEIN EXPRESSION USING DOUBLE-STRANDED CONCATEMERIC DNA INCLUDING PHOSPHOROTHIOATED NUCLEOTIDES

IN VIVO RNA OR PROTEIN EXPRESSION USING DOUBLE-STRANDED CONCATEMERIC DNA INCLUDING PHOSPHOROTHIOATED NUCLEOTIDES

机译：含磷酸化核苷酸的双链消费DNA在体内RNA或蛋白质表达

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摘要

A method for in vivo RNA or protein expression is provided. The method includes introducing a double-stranded concatemeric DNA into a eukaryotic cell to generate a desired RNA or protein. The double-stranded concatemeric DNA includes a plurality of tandem repeat sequences, wherein each of the plurality of tandem repeat sequences comprises an expression sequence. The double-stranded concatemeric DNA comprises one or more phosphorothioated nucleotides, wherein a ratio of phosphorothioated nucleotides to total nucleotides in the double-stranded concatemeric DNA is at least 1:1600. A eukaryotic cell comprising an exogeneous, double-stranded concatemeric DNA comprising the plurality of tandem repeat sequences is also provided.

机译：提供了一种用于体内 RNA或蛋白质表达的方法。该方法包括将双链串联体DNA引入真核细胞以产生所需的RNA或蛋白质。双链串联体DNA包括多个串联重复序列，其中多个串联重复序列中的每一个都包含表达序列。双链串联体DNA包含一个或多个硫代磷酸酯化核苷酸，其中双链串联体DNA中硫代磷酸酯化核苷酸与总核苷酸的比率为至少1：1600。还提供了包含含有多个串联重复序列的外源双链串联DNA的真核细胞。 展开▼

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公开/公告号WO2019053039A1

专利类型

公开/公告日2019-03-21

原文格式PDF

申请/专利权人 GENERAL ELECTRIC COMPANY;GE HEALTHCARE BIO-SCIENCES AB;
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申请/专利号WO2018EP74559

发明设计人 DAVIS BRIAN MICHAEL;NELSON JOHN RICHARD;GAO WEI;
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申请日2018-09-12

分类号C12N15/63;C12N15/67;

国家 WO

入库时间 2022-08-21 11:55:53

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1. 使用包括硫代磷酸酯化的核苷酸的双链多联体DNA的体内RNA或蛋白质表达 [P] . 中国专利： CN111094574A . 2020-05-01

2. 使用双链多联体DNA的无细胞蛋白质表达 [P] . 中国专利： CN110997922A . 2020-04-10

3. VIVO RNA OR PROTEIN EXPRESSION USING DOUBLE-STRANDED CONCATEMERIC DNA INCLUDING PHOSPHOROTHIOATED NUCLEOTIDES [P] . 美国专利： US2019083655A1 . 2019-03-21

机译：使用含磷脂酰化核苷酸的双链消费DNA进行体内RNA或蛋白质表达

4. In vivo RNA or protein expression using double-stranded concatemeric DNA including phosphorothioated nucleotides [P] . 美国专利： US10350307B2 . 2019-07-16

机译：使用包括硫代磷酸化核苷酸的双链串联DNA进行体内RNA或蛋白质表达

5. RNA or protein expression in vivo using double-stranded concatemer DNA containing phosphorothioated nucleotides [P] . 韩国专利： KR20200051648A . 2020-05-13

机译：使用含硫代磷酸核苷酸的双链串联体DNA在体内表达RNA或蛋白质

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11. Nucleotide Sequence of the Hantaan Virus S RNA Segment and Expression of Encoded Proteins. [R] . Jennings, G. B. 1987

1. BACKGROUND： Vibrio vulnifi cus inside the body could activate the NF-!B signaling pathwayand initiate the inflammatory cascade. The lung is one of the earliest organs affected by sepsisassociated with acute lung injury. High mobility group protein B1 （HMGB1） is an important late-actingpro-infl ammatory cytokine involving in the pathophysiology of sepsis. It is also involved in the injuryprocess in the lung, liver and intestine. There has been no report on the involvement of HMGB1 inVibrio vulnifi cus sepsis-induced lung injury.METHODS： Sixty rats were randomly divided into a normal control group （group A, n=10） anda Vibrio vulnificus sepsis group （group B, n=50）. Sepsis was induced in the rats by subcutaneousinjection of Vibrio vulnificus （concentration 6×108 cfu/mL, volume 0.1 mL/100g）） into the left lowerlimbs. The rats in group B were sacrifi ced separately 1, 6, 12, 24, and 48 hours after the infection.Their lungs were stored as specimens, lung water content was measured, and lung pathology wasobserved under a light microscope. The expressions of the HMGB1 gene and protein in the lungswere detected by RT-PCR and Western blot. Data were analyzed with one-way analysis of variance（ANOVA） and the LSD method for pair-wise comparison between the two groups. P〈0.05 wasconsidered statistically signifi cant.RESULTS： Compared to group A （0.652±0.177）, HMGB1 mRNA expression in the lungs ofgroup B was signifi cantly higher at 0 hour （1.161±0.358, P=0.013）, 24 hours （1.679±0.235, P=0.000）,and 48 hours （1.258±0.274, P=0.004）（P〈0.05）, and peaked at 24 hours. Compared to group A（0.594±0.190）, HMGB1 protein expression at 6 hours （1.408±0.567, P=0.026） after infection wassignificantly increased （P〈0. 05）, and peaked at 24 hours （2.415±1.064, P=0.000） after infection.Compared to group A （0.699±0.054）, lung water content was significantly increased at 6 hours（0.759±0.030, P=0.001）,12 hours （0.767±0.023, P=0.000）, 24 hours （0.771±0.043, P=0.000） and 48hours （0.789±0.137, P=0.000） after infection （P〈0.05）. Compared to group A, pathological changesat 12 hours in group B indicate marked pulmonary vascular congestion, interstitial edema andinfl ammatory infi ltration. Alveolar cavity collapse and boundaries of the alveolar septum could not beclearly identifi ed.CONCLUSION： Vibrio vulnifi cus sepsis can lead to injury in rat lungs, and increased HMGB1expression in lung tissue may be one of the mechanisms for injury from Vibrio vulnifi cus sepsis. [J] . Qiao-meng Qiu ,Zhong-wang Li ,Lu-ming Tang . 世界急诊医学杂志(英文) . 2011,第4期

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4. 具双T-DNA区的大麦黄矮病毒复制酶基因双链RNAi植物表达载体构建 [A] . 孙文瑜 . 2009