哺乳动物中RNA干扰研究进展

RNA干扰(RNAi)由双链RNA引起,广泛存在于动、植物中的序列特异性转录后基因沉默,是生物体在进化过程中,抵御病毒感染及由于重复序列和突变引起的基因组不稳定性的保护机制。论文从RNAi现象、RNAi发生机制、哺乳动物中的RNAi现象和RNAi与抗病毒感染等几个方面做了综述,详细介绍RNAi在哺乳动物中的应用。     

RNA干扰技术在小麦中的应用研究进展

小麦是六倍体,其染色体组中存在着大量的同源基因,其育种的主要目标是获得可遗传的优良性状.RNA干扰(RNAi)是指由双链RNA介导的转录后基因沉默.该技术具有两大特点:一是可同时沉默多基因家族及多倍体中的同源基因;二是其沉默效应可在后代中稳定遗传.这使得RNAi在小麦功能基因组学和品质改良研究中的应用具有独特优势.本文对RNAi在小麦功能基因组学和品质改良研究中的应用及研究进展进行了总结,指出了目前存在的问题,并阐明了RNAi在小麦研究中的应用前景.     

RNA干扰技术及其在寄生线虫基因功能研究中的应用

介绍了RNA干扰（RNAi）的概念、线虫中RNAi的作用机理、RNAi的三个分子特性，即可遗传性、转移性RNAi的放大效应和RNAi信号的传播特性。从模板dsRNA的获取、dsRNA的导入和线虫中RNAi效应的检测方法三个方面概述了应用RNAi技术研究线虫基因功能的试验方法和最新进展。     

RNA干涉技术

综述RNAi的发展历史,介绍RNA的作用机制及其特点并展望其应用前景。     

软体动物毛蚶RNA干扰技术的应用研究

为了在软体动物中建立RNA干扰技术,以用于毛蚶基因功能研究。通过体外转录技术合成双链RNA,并将其注射毛蚶干扰目标基因的表达。肌肉注射双链RNA48h后取样,用RT-PCR技术检测目标基因的表达变化,以确定干扰的有效性。结果显示:各目标基因的表达均能被各自特异的双链RNA有效干扰而降低。在注射双链RNA后不同时间点,即12、48、72、96、120h,取样检测目标基因的时序表达,以确定目标基因表达被干扰的时间效应。发现目标基因的表达在48h即能有效降低,而且有效时间能持续至120h以上。对注射双链RNA的动物个体,于48h取样检测不同组织的目标基因表达,发现毛蚶的RNA干扰效应能系统性地在不同组织发生,表明干扰效应能在不同组织之间进行传递。因此,本研究报道了一种稳定的毛蚶RNA干扰技术,为在毛蚶中进行基因功能鉴定和抗病毒研究提供了一个有效的分子生物学的方法。     

RNAi-基因沉默的分子机理

本文介绍了RNA干扰造成基因沉默的几种小分子RNA的类型和作用机理，以及RNA干扰的应用前景及不同方法合成的小分子RNA对动物机体免疫系统的影响。     

Grain Quality as Affected by Down-regulation of Expression of Different ALK Alleles in indica Rice (Oryza sativa L.)

【Objective】Gelatinization temperature (GT) is one of the key physicochemical properties in rice quality, which is mainly regulated by ALK (SSII-3) gene. In general, there are two ALK alleles among indica cultivars. To detect their functional differentiation in indica rice,【Method】Zhenshan 97B (a high GT variety carrying ALKc allele) and Longtefu B (a low GT variety carrying ALKb allele), were used as receptors for the generation of transgenic rice with down-regulation of ALK expression by RNA interference (RNAi).【Result】Down-regulation of ALK gene significantly decreased the GT of the transgenic lines. Due to the difference of original GT between the two receptors, the GT of transgenic rice lines derived from Zhenshan 97B (a high GT variety) decreased significantly, but it is slightly decreased in transgenic plants derived from Longtefu B (a low GT variety). The differential scanning calorimetry (DSC) results showed that the initial temperature of RNAi transgenic rice was significantly lower than the corresponding control and the transgenic lines were gelatinized in advance. The peak value of GT(Tp) in RNAi rice grains was significantly lower than that of the control under Zhenshan 97B background. However, Tp of RNAi rice grains under Longtefu B background was significantly lower than the control to a lesser extent. Also, down-regulation of ALK expression had a significant effect on rice physical-chemical characteristics. An increase of apparent amylose content in RNAi transgenic plants was detected due to the decreased expression of ALK gene. Besides, the pasting properties showed that down-regulation of ALK gene had obvious effects on peak viscosity and breakdown value, improving the taste of the transgenic rice. The gel consistency was significantly different among Zhenshan 97B RNAi lines and their parents, but no difference was found in Longtefu-derived transgenic lines.【Conclusion】RNA interference to ALK allele expression had a significant effect on rice quality, especially the gelatinization characters. Down-regulated expression level of ALKc allele would cause larger variation of physical-chemical characteristics between transgenic rice and their parent than that of ALKb allele.