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Rattler plating beads

Company: ZYMO RESEARCH
Catalog#: S1001
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RNA ImmunoGenic Assay: A Method to Detect Immunogenicity of in vitro Transcribed mRNA in Human Whole Blood
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Date:
2020-12-20
[Abstract]  The mRNA therapeutics is a new class of medicine to treat many various diseases. However, in vitro transcribed (IVT) mRNA triggers immune responses due to recognition by human endosomal and cytoplasmic RNA sensors, but incorporation of modified nucleosides have been shown to reduce such responses. Therefore, an assay signifying important aspects of the human immune system is still required. Here, we present a simple ex vivo method called ‘RNA ImmunoGenic Assay’ to measure immunogenicity of IVT-mRNAs in human whole blood. Chemically modified and unmodified mRNA are complexed with a transfection reagent (TransIT), and co-incubated in human whole blood. Specific cytokines are measured (TNF-α, INF-α, INF-γ, IL-6 and IL-12p70) using ELISAs. The ... [摘要]  [摘要] mRNA疗法是治疗多种疾病的新型药物。然而,我Ñ体外转录(IVT)的mRNA触发由于人类内体和细胞质RNA传感器,但是修饰的核苷的掺入识别的免疫应答已经示出吨ö减少此类反应。牛逼herefore ,测定标志着重要的环节人体免疫系统仍然需要。这里,我们提出一个简单的离体称为“RNA方法免疫ģ ENIC测定”测量的IVT-mRNA的免疫原性小号在人全血。将化学修饰和未修饰的mRNA与转染试剂(TransIT )复合,并在人全血中共同孵育。特异性细胞因子测定(TNF- α ,INF- α ,INF- γ ,IL-6和IL-12p70的),使用的ELISA。进行qPCR分析以揭示特异性免疫途径的激活。所述RNA免疫ģ ENIC测定提供小号的简单且快速的方法来检测供体特异性-针对mRNA的治疗剂的免疫应答。

图形摘要:

RNA免疫基因测定的示意图

[背景] mRNA治疗是基因治疗的重要一类(Sahin等,2014 ;Antony等,2015 ...

Ty1 Retrotransposition Frequency Assay Using a Chromosomal Ty1his3AI or Ty1kanMXAI Element
Author:
Date:
2018-09-05
[Abstract]  Here I describe a simple genetic assay to determine the frequency of retrotransposition of a single chromosomal Ty1 element that is marked with the retrotransposition indicator gene, his3AI or kanMXAI. The assay is used to determine the effect of mutations or environmental conditions on the frequency of Ty1 retrotransposition in the yeast, Saccharomyces cerevisiae. [摘要]  在这里,我描述了一个简单的基因测定,以确定单个染色体Ty1元件的逆转录频率,该元件用逆转录指示基因 his3AI 或 kanMXAI 标记。 该测定用于确定突变或环境条件对酵母中的Ty1逆转录频率的影响, Saccharomyces cerevisiae 。

【背景】Ty1是长末端重复(LTR)反转录转座子,其在结构上和进化上与逆转录病毒相关。当Ty1 RNA在细胞质病毒样颗粒内逆转录时,Ty1的逆转录发生,导致cDNA的合成,转运回细胞核并整合到其宿主的基因组中(Boeke et al。 ,1985; Garfinkel et al。,1985)。整合的Ty1元件包含两个相同方向的LTR,位于两个开放阅读框的侧翼: GAG ,它编码形成病毒样颗粒并结合Ty1 RNA的结构蛋白, POL ,它编码三种酶蛋白 - 蛋白酶,逆转录酶和整合酶(图1)。 Ty1元素是 S中所有五个LTR-反转录转座子家族中最活跃和最丰富的转录。酵母。 Ty1已被证明受数百种基因,几种不同的环境条件和各种DNA损伤剂的调节(Curcio et ...

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