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Rapid and accurate species identification for ecological studies and monitoring using CRISPR-based SHERLOCK.
Molecular Ecology Resources ( IF 5.5 ) Pub Date : 2020-05-12 , DOI: 10.1111/1755-0998.13186
Melinda R Baerwald 1 , Alisha M Goodbla 2 , Raman P Nagarajan 2 , Jonathan S Gootenberg 3, 4, 5, 6, 7 , Omar O Abudayyeh 3, 4, 5, 6, 8 , Feng Zhang 3, 4, 5, 6 , Andrea D Schreier 2
Affiliation  

One of the most fundamental aspects of ecological research and monitoring is accurate species identification, but cryptic speciation and observer error can confound phenotype‐based identification. The CRISPR‐Cas toolkit has facilitated remarkable advances in many scientific disciplines, but the fields of ecology and conservation biology have yet to fully embrace this powerful technology. The recently developed CRISPR‐Cas13a platform SHERLOCK (Specific High‐sensitivity Enzymatic Reporter unLOCKing) enables highly accurate taxonomic identification and has all the characteristics needed to transition to ecological and environmental disciplines. Here we conducted a series of “proof of principle” experiments to characterize SHERLOCK’s ability to accurately, sensitively and rapidly distinguish three fish species of management interest co‐occurring in the San Francisco Estuary that are easily misidentified in the field. We improved SHERLOCK’s ease of field deployment by combining the previously demonstrated rapid isothermal amplification and CRISPR genetic identification with a minimally invasive and extraction‐free DNA collection protocol, as well as the option of instrument‐free lateral flow detection. This approach opens the door for redefining how, where and by whom genetic identifications occur in the future.

中文翻译:

使用基于CRISPR的SHERLOCK对生态研究和监测进行快速准确的物种鉴定。

准确的物种识别是生态学研究和监测的最基本方面之一,但是神秘的物种形成和观察者错误会混淆基于表型的识别。CRISPR‐Cas工具箱推动了许多科学领域的显着进步,但生态学和保护生物学领域尚未完全接受这项强大的技术。最近开发的CRISPR‐Cas13a平台SHERLOCK(特定的高灵敏度酶报告基因解锁)可进行高度准确的分类识别,并具有过渡到生态和环境学科所需的所有特征。在这里,我们进行了一系列的“原理验证”实验,以表征SHERLOCK准确,灵敏快速地区分在旧金山河口同时出现的三种具有管理意义的鱼类,这些鱼类很容易在野外被错误识别。我们将先前证明的快速等温扩增和CRISPR遗传学鉴定与微创,无提取DNA收集方案以及无仪器侧向流检测选项相结合,提高了SHERLOCK的现场部署便利性。这种方法为重新定义将来如何,在何处以及由谁进行遗传鉴定打开了大门。以及可选的无仪器侧向流量检测。这种方法为重新定义将来如何,在何处以及由谁进行遗传鉴定打开了大门。以及可选的无仪器侧向流量检测。这种方法为重新定义将来如何,在何处以及由谁进行遗传鉴定打开了大门。
更新日期:2020-05-12
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