Enzymatic Tools for Degrading Tissue and Preserving RNA

描述(由申请人提供)： 完整的RNA是表达谱研究的必要起始材料 可以将特定分子事件与疾病表型联系起来的技术。 从组织中分离RNA目前需要侵入性机械力来 颠覆蜂窝架构。这种机械的方法是枯燥乏味的 潜在的低效。它们还使操作员暴露在可能的生物危害中。 因此，有必要开发一种用于RNA分离的“封闭系统” 为了能够高通量地从组织(如活检)中回收RNA 使用一个受保护的平台，可以在 一次。我们建议通过使用“熔化”来实现这一目标。 (组织多酶液化)技术。这项技术将 利用酶的降解能力，这些酶作为一种 用于分解复杂食物来源的生物工具集。我们指的是 这个工具集成了一个“酵素社区”。因为一些酶群落(例如， 细菌病原体的分泌酶)模拟了活动的光谱 需要消化组织，这些收集是一个理想的存储库 组织降解酶。我们建议对酶进行表征和混合 产生一种能迅速降解组织的熔融剂的活动，“冷冻” MRNA表达谱，并稳定RNA用于下游分析。

DESCRIPTION (provided by applicant): Intact RNA is an obligate starting material for expression profiling technologies that can link specific molecular events with disease phenotypes. RNA isolations from tissue currently require invasive mechanical force to disrupt the cellular architecture. Such mechanical approaches are tedious and potentially inefficient. They also expose the operator to possible biohazards. As a result, the development of a "closed system" for RNA isolation is needed to enable the high-throughput recovery of RNA from tissues such as biopsies using a safeguarded platform that can process dozens or hundreds of tissues at once. We proposed to achieve this goal through the use of "MELT" (Multi-Enzymatic Liquefaction of Tissue) technology. This technology will harness the degrading power of enzymes that have evolved together as a biological tool set for the decomposition of complex food sources. We refer to this tool set as an "enzyme community." Since some enzyme communities (e.g., the secreted enzymes of bacterial pathogens) mimic the spectrum of activities needed to digest tissue, these collections are an ideal repository for tissue-degrading enzymes. We propose to characterize and blend enzyme activities to create a MELT reagent that can rapidly degrade tissues, "freeze" mRNA expression profiles, and stabilize the RNA for downstream analyses.