CRISPR/dCas9-assisted SERS sensing for transgenic crops based on two-factor verification of 35S promoter and Nos terminator

文献类型： 外文期刊

第一作者： Su, Ailing

作者： Su, Ailing;Xu, Weiqing;Xu, Shuping;Chen, Ziqi;Liu, Xiangguo;Liang, Chongyang;Xu, Shuping;Xu, Shuping

作者机构：

关键词： CRISPR/dCas9; SERS; Gene detection; Transgenic crop identification; Multiplex detection

期刊名称：SENSORS AND ACTUATORS B-CHEMICAL （ 影响因子：7.7； 五年影响因子：7.4 ）

ISSN：

年卷期： 2025 年 429 卷

页码：

收录情况： SCI

摘要： A nucleic acid analysis platform was built based on the combination of the recombinase polymerase amplification (RPA)-aided CRISPR/dCas9 and surface-enhanced Raman scattering (SERS) technologies, which enables the simultaneous detection of dual nucleic acid fragments, for the purpose of identifying transgenic crops. Two SERS-active nanoprobes (AgNP@Ra@dCas9/sgRNA) were fabricated by grafting the single RNA (sgRNA)-activated dCas9 protein to silver nanoparticles that have been decorated with two different Raman reporters (Ra). RPA primers endow target genes with biotin end that can be captured by the streptavidin-modified magnetic bead (MB@SA), followed by the conjugation of SERS nanoprobes. Thus, strong SERS signals would be detected above the MBs. This SERS platform has been applied for identifying transgenic crops based on the sequence designs specifically for the 35S promoter and Nos terminator simultaneously. Double confirmation of target genes can ensure the accuracy of detection. The whole sensing process, including RPA, dCas9 activation, and target gene identification, can be completed in one tube, within 80 min. This CRISPR/dCas9-SERS analytical platform has been proved to be feasible in detecting plant samples, indicating one successful attempt of the CRISPR/dCas9-SERS systems in multi-target detection.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序