Harnessing intermolecular G-quadruplex-based spatial confinement effect for accelerated activation of CRISPRCas12a empowers ultra-sensitive detection of PMLRARA fusion genes
Harnessing intermolecular G-quadruplex-based spatial confinement effect for accelerated activation of CRISPRCas12a empowers ultra-sensitive detection of PMLRARA fusion genes
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DOI：
10.1016/j.aca.2023.342108
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发表时间：
2023
影响因子：
6.2
通讯作者：
Juan Xia
中科院分区：
文献类型：
--
作者：
Xinrui Wang;Dan Zheng;Chengyi Wang;Danni Xue;Qi Wang;Juan Xia
Accurate detection and classification of the three isoforms of PML/RARA genomic fragments are crucial for predicting disease progression, stratifying risk, and administering precise drug therapies in acute promyelocytic leukemia (APL). In this study, we have developed a highly specific nucleic acid detection platform capable of quantifying the long isoform of the three main PML-RARA isoforms at a constant temperature. This platform integrates the strengths of the CRISPR/Cas12a nuclease-based method and the rolling circle amplification (RCA) technique. Notably, the RCA-assisted CRISPR/Cas12a trans-cleavage system incorporates a spatial confinement effect by utilizing intermolecular G-quadruplex structures. This innovative design effectively enhances the local concentration of CRISPR/Cas12a, thereby accelerating its cleaving efficiency towards reporter nucleic acids and enabling the detection of PML/RARA fusion gene expression through spectroscopy. The robust detection of PML/RARA fusion gene from human serum samples validates the reliability and potential of this platform in the screening, diagnosis, and prognosis of APL cases. Our findings present an approach that holds significant potential for the further development of the robust CRISPR/Cas sensor system, offering a rapid and adaptable paradigm for APL diagnosis.
