CRISPR-induced DNA reorganization for multiplexed nucleic acid detection.

IF 14.7 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Nature Communications Pub Date : 2023-03-17 DOI:10.1038/s41467-023-36874-6

Margot Karlikow, Evan Amalfitano, Xiaolong Yang, Jennifer Doucet, Abigail Chapman, Peivand Sadat Mousavi, Paige Homme, Polina Sutyrina, Winston Chan, Sofia Lemak, Alexander F Yakunin, Adam G Dolezal, Shana Kelley, Leonard J Foster, Brock A Harpur, Keith Pardee

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引用次数: 7

Abstract

Nucleic acid sensing powered by the sequence recognition of CRIPSR technologies has enabled major advancement toward rapid, accurate and deployable diagnostics. While exciting, there are still many challenges facing their practical implementation, such as the widespread need for a PAM sequence in the targeted nucleic acid, labile RNA inputs, and limited multiplexing. Here we report FACT (Functionalized Amplification CRISPR Tracing), a CRISPR-based nucleic acid barcoding technology compatible with Cas12a and Cas13a, enabling diagnostic outputs based on cis- and trans-cleavage from any sequence. Furthermore, we link the activation of CRISPR-Cas12a to the expression of proteins through a Reprogrammable PAIRing system (RePAIR). We then combine FACT and RePAIR to create FACTOR (FACT on RePAIR), a CRISPR-based diagnostic, that we use to detect infectious disease in an agricultural use case: honey bee viral infection. With high specificity and accuracy, we demonstrate the potential of FACTOR to be applied to the sensing of any nucleic acid of interest.

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CRISPR诱导DNA重组用于多重核酸检测。

由CRIPSR技术的序列识别提供动力的核酸传感使快速、准确和可部署的诊断取得了重大进展。尽管令人兴奋，但它们的实际实施仍面临许多挑战，例如在靶向核酸中广泛需要PAM序列、不稳定的RNA输入和有限的多路复用。在这里，我们报道了FACT（功能化扩增CRISPR追踪），这是一种基于CRISPR的核酸条形码技术，与Cas12a和Cas13a兼容，能够基于任何序列的顺式和反式切割进行诊断输出。此外，我们通过可再编程PAIRing系统（RePAIR）将CRISPR-Cas12a的激活与蛋白质的表达联系起来。然后，我们将FACT和RePAIR结合起来创建FACTOR（FACT on RePAIR），这是一种基于CRISPR的诊断方法，用于检测农业用例中的传染病：蜜蜂病毒感染。凭借高特异性和准确性，我们证明了FACTOR应用于感兴趣的任何核酸的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nature Communications Biological Science Disciplines-

CiteScore

24.90

自引率

2.40%

发文量

6928

审稿时长

3.7 months

期刊介绍： Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.