정주연 (한국생명공학연구원)

한빛사논문

조회304

한국생명공학연구원

Biosens. Bioelectron., Available online 22 October 2022 | https://doi.org/10.1016/j.bios.2022.114819 Hybrid CRISPR/Cas protein for one-pot detection of DNA and RNA

Kyeonghye Guk^a,1, Soyeon Yi^a,¹, Hyeran Kim^a, Yoonji Bae^a, Dongeun Yong^b, Sunjoo Kim^c, Kyu-Sun Lee^a,e, Eun-Kyung Lim^a,d,e, Taejoon Kang^a,e, Juyeon Jung^a,d,e

^aBionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea
^bDepartment of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University, 50-1 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea
^cRepublic of Korea Department of Laboratory Medicine, Gyeongsang National University College of Medicine, Jinju, 52727, Republic of Korea
^dDepartment of Nanobiotechnology, KRIBB School of Biotechnology, UST, 217 Gajeong-ro, Yuseong-gu, Daejeon, 34113, Republic of Korea
^eSchool of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 16419, Republic of Korea

¹These authors equally contributed to this work.
Corresponding authors: Taejoon Kang, Juyeon Jung

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostics have emerged as next-generation molecular diagnostics. In CRISPR-based diagnostics, Cas12 and Cas13 proteins have been widely employed to detect DNA and RNA, respectively. Herein, we developed a novel hybrid Cas protein capable of detecting universal nucleic acids (DNA and RNA). The CRISPR/hybrid Cas system simultaneously recognizes both DNA and RNA, enabling the dual detection of pathogenic viruses in a single tube. Using wild-type (WT) and N501Y mutant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as detection models, we successfully detected both virus strains with a detection limit of 10 viral copies per reaction without cross-reactivity. Furthermore, it is demonstrated the detection of WT SARS-CoV-2 and N501Y mutant variants in clinical samples by using the CRISPR/hybrid Cas system. The hybrid Cas protein is expected to be utilized in a molecular diagnostic method for infectious diseases, tissue and liquid biopsies, and other nucleic acid biomarkers.

논문정보

형식Research article
게재일2022년 10월 (BRIC 등록일 2022-11-22)
연구진국내 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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