您好，欢迎访问浙江省农业科学院 机构知识库！
浙江省农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Qualitative and Quantitative Detection of CRISPR-Associated Cas Gene in Gene-Edited Foods

文献类型： 外文期刊

作者： Ding, Lin 1 ; Xu, Xiaoli 1 ; Wang, Xiaofu 1 ; Chen, Xiaoyun 1 ; Lu, Yuwen 2 ; Xu, Junfeng 1 ; Peng, Cheng 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, State Key Lab Managing Biot & Chem Threats Qual &, Key Lab Traceabil Agr Genet Modified Organisms, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China

2.Ningbo Univ, Inst Plant Virol, State Key Lab Managing Biot & Chem Threats Qual &, Key Lab Biotechnol Plant Protect MARA & Zhejiang P, Ningbo 315211, Peoples R China

关键词： gene editing technology; gene-edited foods; qualitative PCR; qPCR; Cpf1

期刊名称：FOODS （ 影响因子：5.2； 五年影响因子：5.5 ）

ISSN：

年卷期： 2023 年 12 卷 19 期

页码：

收录情况： SCI

摘要： Effective regulation of gene-edited products and resolution of public concerns are the prerequisites for the industrialization of gene-edited crops and their derived foods. CRISPR-associated protein, the core element of the CRISPR system, requires to be regulated. Thus, there is an urgent need to establish qualitative and quantitative detection methods for the Cas gene. In the present study, the primers and probes were designed and screened for Cas12a (Cpf1), which is the most commonly used target site in gene editing; we performed PCR system optimization, determined the optimal primer concentration and annealing temperature, and established qualitative PCR and quantitative PCR (qPCR) assays for detecting Cpf1 in gene editing by specificity and sensitivity tests. In specificity testing, qualitative PCR and qPCR methods could 100% detect samples containing Cpf1 DNA, while the detection rate of other samples without Cpf1 was 0%. In the assay sensitivity test, the limit of detection of qualitative PCR was 0.1% (approximately 44 copies), and the limit of detection of the qPCR method was 14 copies. In the stability test, both the qualitative PCR and qPCR methods were repeated 60 times at their corresponding lowest detection limit concentrations, and the results were positive. Thus, the qualitative and quantitative assays for Cpf1 are specific, sensitive, and stable. The method provides technical support for the effective monitoring of gene-edited products and their derived foods in the future.

  • 相关文献
作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：浙江省农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台