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题名	一种新型聚苯乙烯基质RNA固相有机合成载体制备的研究
作者	梁洪
学位类别	硕士
答辩日期	2009-05-26
授予单位	中国科学院过程工程研究所
授予地点	过程工程研究所
导师	苏志国
关键词	siRNA RNA固相有机合成 固相载体 聚苯乙烯 通用间臂
其他题名	Study on a new universal polystyrene support for solid phase organic synthesis of RNA
学位专业	生物化工
中文摘要	小分子干扰RNA(small interfering RNA, siRNA)是一种21-23bp的双链RNA，它能引起RNA干扰(RNA interference, RNAi)，在mRNA水平关闭相应序列基因表达或使其沉默，在抗病毒、抗肿瘤和心血管疾病治疗等方面具有很大的潜力。固相有机合成是目前获得大量siRNA最有效的途径。现已上市的固相有机合成载体多以CPG为基质，在RNA合成过程中，传统的CPG载体存在介质表面亲水导致彻底除水困难影响偶联效率、产生错误序列、载体不能满足各种RNA序列合成需要等缺点，对RNA合成带来了一定的影响。开发一种新型的RNA固相有机合成载体，是解决上述问题的有效途径之一。 针对目前上市的CPG载体存在表面亲水、不具备通用性等缺点，本研究将氨甲基衍生的聚苯乙烯载体通过琥珀酸键与3-氨基-1-(4,4’-二甲氧三苯甲基)-1,2-丙二醇偶联，得到载量更高、合成性能更好、应用范围广、具有自主知识产权的RNA固相有机合成载体。 本论文以提高反应效率和载体性能为目标，对载体合成过程中聚苯乙烯的氨甲基衍生、3-氨基-1-(4,4’-二甲氧三苯甲基)-1,2-丙二醇间臂的合成及其与载体的偶联过程中各影响因素，如原料投料比、催化剂、反应时间、反应温度、分离纯化方法等分别进行了考察，优化出最佳反应条件，建立了高效、低成本的载体制备工艺。 本论文还对自制的聚苯乙烯RNA固相有机合成载体的性能进行了考察，载体应用范围广，能够满足不同RNA序列的合成需要；有效载量达61-92 µmol/g，为传统载体载量的2倍左右；偶联效率大于95%，与传统载体类似；载体合成产物与传统载体合成产物具有同一性。因此，这是一种有潜力替代传统CPG载体的新型聚苯乙烯基质RNA固相有机合成载体。
英文摘要	Small interfering RNA (siRNA) is 21-23bp double-stranded RNA, which can direct a sequence-specific post-transriptional gene silencing, a process commonly referred to as RNA interference or RNAi. Because of this property, siRNA has the potential to be used as anti-virus and anti-tumor drugs. So far, the most effective way to obtain siRNA is solid phase organic synthesis. Controlled pore glass (CPG) is the most commonly used solid phase support, but its hydrophilic surface could render the coupling reaction inefficient and produce mis-matched sequence. Such drawbacks limit its ues in synthesis of different RNA products. To solve these problems, polystyrene was chosen to synthesize a new type of solid phase support for RNA synthesis in this study. This new type of solid phase support was prepared through attachment of 3-amino-1-(4,4’-dimethoxytriphenylmethyl)-2-propa- nediol linker toaminomethyl-polystyrene support via succinic anhydride. Compared with the conventional CPG support, the polystyrene support can be used to synthesize different RNA sequence, with high loading capacities. The effect of each reaction condition on the performance of the support and the loading was studied. An optimized synthesis process with low costs and high efficiency was established. The performance of the new support was compared with that of the conventional CPG. The synthetic efficiencies were almost at the same level. However, the loading of activation site of the new support was 61-92µmol/g, which was much higher than that of CPG. Moreover, the new support could be used as a universal support to synthesize any sequence RNA. RP-HPLC analysis showed that the products synthesized on the universal polystyrene support and on the CPG support were almost identical. Therefore, this universal solid matrix possessed potential to replace the conventional CPG support in RNA synthesis.
语种	中文
公开日期	2013-09-16
页码	84
内容类型	学位论文
源URL	[http://ir.ipe.ac.cn/handle/122111/1290]
专题	过程工程研究所_研究所（批量导入）
推荐引用方式 GB/T 7714	梁洪. 一种新型聚苯乙烯基质RNA固相有机合成载体制备的研究[D]. 过程工程研究所. 中国科学院过程工程研究所. 2009.

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