高级检索
当前位置: 首页 > 详情页

Sensitive and reliable miRNA analysis based on cyclic reverse transcription and CRISPR-Cas12a-assisted signal cycle

文献详情

资源类型:
WOS体系:

收录情况: ◇ SCIE

机构: [1] Obstetrics Department, Shijiazhuang Fourth Hospital, Hebei Medical University, Shijiazhuang City 050000, Hebei Province, China [2] Physical Examination Center, Shijiazhuang Fourth Hospital, Hebei Medical University, Shijiazhuang City 050000, Hebei Province, China
出处:
ISSN:

关键词: MicroRNAs (miRNAs) Cyclic reverse transcription (CRT) CRISPR-Cas12a Duplex-specific nuclease (DSN)

摘要:
MicroRNAs (miRNAs), a category of small molecules that possess significant regulatory capabilities, have been extensively employed as biomarkers in the domain of biosensing to facilitate the early detection of diverse ailments. However, sensitive and accurate miRNA detection remains a huge challenge due to the high similarity between the homologous sequences and low abundance. Therefore, it is essential to develop methods with high sensitivity and specificity for miRNA detection. In this study, we present the development of a signal cycle-based platform that utilizes cyclic reverse transcription (CRT) and CRISPR-Cas12a to enable the precise and sensitive detection of microRNAs. The CRT mechanism facilitates precise target recognition in the presence of target miRNA, thereby converting miRNA signals to DNA signals. The trans-cleavage activity of the Cas12a protein is triggered by the formation of complete hairpin-shaped CRT products; this results in the cleavage of the DNA section contained in the H probe, while the RNA section ("4"@MBs) remains loaded onto the surface of magnetic beads (MB). By binding with the "reporter" sensor, the "4" sequences create an RNA/DNA duplex that the duplex-specific nuclease (DSN) can recognize. The "reporter" probe is thus metabolized, leading to the reappearance of the fluorescence signal. By capitalizing on the exceptional fidelity and selectivity of CRISPR/Cas12a, as well as the substantial impact of triggered enzymatic cycle amplification, this approach demonstrated remarkable sensitivity and specificity in miRNA detection, even in a complex environment containing 10% fetal bovine serum (FBS) and a serum sample. In contrast, a detection limit of 3.2 fM is conceivable. Furthermore, this approach maintained a notable degree of stability, which was anticipated to result in the detection of miRNAs in an effective and sensitive manner.

基金:
语种:
WOS:
中科院分区:
出版当年[2025]版:
大类 | 3 区 化学
小类 | 3 区 分析化学
最新[2025]版:
大类 | 3 区 化学
小类 | 3 区 分析化学
JCR分区:
出版当年[2024]版:
Q2 CHEMISTRY, ANALYTICAL
最新[2024]版:
Q2 CHEMISTRY, ANALYTICAL

影响因子: 最新[2024版] 最新五年平均 出版当年[2024版] 出版当年五年平均 出版前一年[2023版]

第一作者:
第一作者机构: [1] Obstetrics Department, Shijiazhuang Fourth Hospital, Hebei Medical University, Shijiazhuang City 050000, Hebei Province, China
共同第一作者:
通讯作者:
推荐引用方式(GB/T 7714):
APA:
MLA:

资源点击量:42329 今日访问量:0 总访问量:1365 更新日期:2025-08-01 建议使用谷歌、火狐浏览器 常见问题

技术支持:重庆聚合科技有限公司 地址:河北省石家庄市健康路12号