Scalable Nucleic Acid Storage and Retrieval Using Barcoded Microcapsules
Scalable Nucleic Acid Storage and Retrieval Using Barcoded Microcapsules
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DOI：
10.1021/acsami.1c14985
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发表时间：
2021-10-15
影响因子：
9.5
通讯作者：
Bathe, Mark
中科院分区：
文献类型：
--
作者：
Banal, James L.;Bathe, Mark
核酸测序和合成技术的快速发展刺激了对病毒、细菌和哺乳动物来源和生物体的DNA和RNA的收集、储存和测序的主要需求。然而，目前储存核酸的方法依赖于低温环境，并且需要机器人才能进入，这对可扩展和低成本的核酸储存提出了挑战。在这里，我们提出了一种储存核酸的替代方法，称为使用条形码微胶囊保存和获取核酸（PANDORA）。从千碱基到千兆碱基的不同来源的核酸，包括动物、细菌和病毒，被封装在二氧化硅微胶囊中，以保护它们在室温下不受环境变性剂的影响。附着在每个微胶囊上的分子条形码使样品池和随后的荧光激活分选识别和检索成为可能。我们演示了定量存储和快速访问目标核酸，模拟了传统存储系统中实施的标准检索操作，包括恢复100,000-200,000个样本和使用四个唯一条形码的布尔逻辑选择。定量聚合酶链反应和短读测序验证了分选实验和释放核酸的完整性。我们提出的方法提供了一种可扩展的长期，室温储存和检索具有高样本保真度的核酸。
Rapid advances in nucleic acid sequencing and synthesis technologies have spurred a major need to collect, store, and sequence the DNA and RNA from viral, bacterial, and mammalian sources and organisms. However, current approaches to storing nucleic acids rely on a low-temperature environment and require robotics for access, posing challenges for scalable and low-cost nucleic acid storage. Here, we present an alternative method for storing nucleic acids, termed Preservation and Access of Nucleic aciDs using barcOded micRocApsules (PANDORA). Nucleic acids spanning kilobases to gigabases and from different sources, including animals, bacteria, and viruses, are encapsulated into silica microcapsules to protect them from environmental denaturants at room temperature. Molecular barcodes attached to each microcapsule enable sample pooling and subsequent identification and retrieval using fluorescence-activated sorting. We demonstrate quantitative storage and rapid access to targeted nucleic acids from a pool emulating standard retrieval operations implemented in conventional storage systems, including recovery of 100,000-200,000 samples and Boolean logic selection using four unique barcodes. Quantitative polymerase chain reaction and short-read sequencing of the retrieved samples validated the sorting experiments and the integrity of the released nucleic acids. Our proposed approach offers a scalable long-term, room-temperature storage and retrieval of nucleic acids with high sample fidelity.