DataVXAdvanced SciencePaper
journal article·2024-11Open access

Unveiling Cas12j Trans‐Cleavage Activity for CRISPR Diagnostics: Application to miRNA Detection in Lung Cancer Diagnosis

Ju‐Eun Kang, Hansol Kim, Young‐Hoon Lee, Ha‐Yeong Lee, Yeonkyung Park, Hyowon Jang, Jae‐Rin Kim, Min‐Young Lee, Byeong‐Ho Jeong, Ju‐Young Byun, Seung Jun Kim, Eun‐Kyung Lim, Juyeon Jung, Eui‐Jeon Woo, Taejoon Kang, Kwang‐Hyun Park

Advanced Science· Volume 11· Issue 44
Publisher pageOpen full text
Source abstract

Abstract

Abstract Cas12j, a hypercompact and efficient Cas protein, has potential for use in CRISPR diagnostics, but has not yet been used because the trans‐cleavage activity of Cas12j is veiled. Here, the trans‐cleavage behavior of Cas12j1, 2, and 3 variants and evaluate their suitability for nucleic acid detection is unveiled. The target preferences and mismatch specificities of the Cas12j variants are precisely investigated and the optimal Cas12j reaction conditions are determined. As a result, the EXP‐J assay for miRNA detection by harnessing the robust trans‐cleavage activity of Cas12j on short ssDNA is developed. The EXP‐J method demonstrates exceptional detection capabilities for miRNAs, proving that Cas12j can be a pivotal component in molecular diagnostics. Furthermore, the translational potential of the EXP‐J assay is validated by detecting oncogenic miRNAs in plasma samples from lung cancer patients. This investigation not only elucidates the trans‐cleavage characteristics of Cas12j variants, but also advances the Cas12j‐based diagnostic toolkit.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: CRISPR and Genetic Engineering · Advanced biosensing and bioanalysis techniques · RNA and protein synthesis mechanisms

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Critical Diseases Diagnostics Convergence Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB) Daejeon 34141 Republic of Korea

Department of Proteome Structural Biology KRIBB School of Bioscience University of Science and Technology (UST) Daejeon 34113 Republic of Korea

Bionanotechnology Research Center KRIBB Daejeon 34141 Republic of Korea

Department of Nano‐Bio Convergence, Surface Materials Division Korea Institute of Materials Science (KIMS) Changwon Gyeongsangnam‐do 51508 Republic of Korea

Division of Pulmonary and Critical Care Medicine Department of Medicine Samsung Medical Center Sungkyunkwan University (SKKU) School of Medicine Seoul 06351 Republic of Korea

School of Pharmacy SKKU Suwon Gyeongi‐do 16419 Republic of Korea

Department of Nanobiotechnology KRIBB School of Biotechnology, UST Daejeon 34113 Republic of Korea

Disease Target Structure Research Center KRIBB Daejeon 34141 Republic of Korea

Funding

Ministry of Agriculture, Food and Rural Affairs · Korea Research Institute of Bioscience and Biotechnology · National NanoFab Center · Korea Evaluation Institute of Industrial Technology · Korea Environmental Industry and Technology Institute

References (59)

Show source-provided references
1. 10.1126/science.add864310.1126/science.add8643 ↗
2. 10.1038/nrmicro356910.1038/nrmicro3569 ↗
3. 10.1038/s41579-019-0299-x10.1038/s41579-019-0299-x ↗
4. 10.1074/jbc.REV120.00703410.1074/jbc.REV120.007034 ↗
5. 10.1038/s41422-018-0022-x10.1038/s41422-018-0022-x ↗
6. 10.1021/acssynbio.9b0020910.1021/acssynbio.9b00209 ↗
7. 10.1126/science.aav429410.1126/science.aav4294 ↗
8. 10.1126/science.aam932110.1126/science.aam9321 ↗
9. 10.1038/s41596-019-0210-210.1038/s41596-019-0210-2 ↗
10. 10.1038/s42003-021-02001-810.1038/s42003-021-02001-8 ↗
11. 10.1038/s41586-020-2279-810.1038/s41586-020-2279-8 ↗
12. 10.1016/j.bios.2022.11481910.1016/j.bios.2022.114819 ↗
13. 10.1016/j.snb.2023.13474810.1016/j.snb.2023.134748 ↗
14. Talwar C. S. · Biosensors (Basel) · 2021 · 11
15. 10.1038/s41467-023-43389-710.1038/s41467-023-43389-7 ↗
16. 10.1016/j.aca.2023.34125810.1016/j.aca.2023.341258 ↗
17. 10.1016/j.cell.2020.12.00110.1016/j.cell.2020.12.001 ↗
18. 10.1016/j.mattod.2022.11.00110.1016/j.mattod.2022.11.001 ↗
19. 10.1016/j.microc.2021.10703610.1016/j.microc.2021.107036 ↗
20. 10.1016/j.tifs.2022.02.03010.1016/j.tifs.2022.02.030 ↗
21. 10.1038/s41551-021-00760-710.1038/s41551-021-00760-7 ↗
22. 10.1126/science.abb140010.1126/science.abb1400 ↗
23. 10.1038/s41594-021-00632-310.1038/s41594-021-00632-3 ↗
24. 10.1038/s41467-021-24707-310.1038/s41467-021-24707-3 ↗
25. 10.1038/s41421-023-00612-510.1038/s41421-023-00612-5 ↗
26. 10.1016/j.tcb.2014.11.00410.1016/j.tcb.2014.11.004 ↗
27. 10.3390/ijms1705062710.3390/ijms17050627 ↗
28. 10.21769/BioProtoc.284210.21769/BioProtoc.2842 ↗
29. 10.1038/s41467-020-18615-110.1038/s41467-020-18615-1 ↗
30. 10.1021/acs.nanolett.1c0071510.1021/acs.nanolett.1c00715 ↗
31. 10.1016/j.bios.2021.11349310.1016/j.bios.2021.113493 ↗
32. 10.1021/acs.analchem.2c0061610.1021/acs.analchem.2c00616 ↗
33. 10.1021/acs.jafc.2c0654910.1021/acs.jafc.2c06549 ↗
34. 10.3390/jcm410189010.3390/jcm4101890 ↗
35. 10.1016/j.bbcan.2010.07.00210.1016/j.bbcan.2010.07.002 ↗
36. 10.1007/s13206-023-00108-010.1007/s13206-023-00108-0 ↗
37. 10.1007/s13206-022-00071-210.1007/s13206-022-00071-2 ↗
38. 10.1038/s41551-023-01033-110.1038/s41551-023-01033-1 ↗
39. 10.1016/j.ymeth.2021.02.01810.1016/j.ymeth.2021.02.018 ↗
40. 10.1039/D1CS00098E10.1039/D1CS00098E ↗
41. 10.1021/acs.analchem.8b0278810.1021/acs.analchem.8b02788 ↗
42. 10.3390/ijms2216866110.3390/ijms22168661 ↗
43. 10.1016/j.arr.2014.02.00910.1016/j.arr.2014.02.009 ↗
44. 10.3389/fmolb.2019.0009810.3389/fmolb.2019.00098 ↗
45. 10.3892/ol.2021.1305310.3892/ol.2021.13053 ↗
46. 10.1007/s13277-013-0753-810.1007/s13277-013-0753-8 ↗
47. 10.3892/ijo.2014.227210.3892/ijo.2014.2272 ↗
48. 10.3390/ijms2204155710.3390/ijms22041557 ↗
49. 10.26717/BJSTR.2020.25.00424110.26717/BJSTR.2020.25.004241 ↗
50. 10.1126/sciadv.abo640510.1126/sciadv.abo6405 ↗
51. 10.1016/j.xplc.2022.10045310.1016/j.xplc.2022.100453 ↗
52. 10.1177/175342591244695510.1177/1753425912446955 ↗
53. 10.1080/10409238.2021.193792710.1080/10409238.2021.1937927 ↗
54. 10.1158/2159-8290.CD-20-131110.1158/2159-8290.CD-20-1311 ↗
55. 10.1016/j.xcrm.2023.10119810.1016/j.xcrm.2023.101198 ↗
56. 10.1016/j.bios.2022.11407110.1016/j.bios.2022.114071 ↗
57. 10.1038/s41467-019-11826-110.1038/s41467-019-11826-1 ↗
58. 10.1038/s41467-021-23497-y10.1038/s41467-021-23497-y ↗
59. 10.1002/smll.20230165410.1002/smll.202301654 ↗