Terminal deoxynucleotidyl transferase induced activators to unlock the trans-cleavage of CRISPR/Cpf 1 (TdT-IU- CRISPR/Cpf 1): An ultrasensitive biosensor for Dam MTase activity detection

被引：34

作者：

Chen, Xiaolong ^{[1
]}

Cao, Gaihua ^{[1
]}

Wang, Xianfeng ^{[1
]}

Ji, Zhong ^{[1
]}

Xu, Faliang ^{[2
]}

Huo, Danqun ^{[1
,3
]}

Luo, Xiaogang ^{[1
]}

Hou, Changjun ^{[1
]}

机构：

[1] Chongqing Univ, Key Lab Biorheol Sci & Technol, State & Local Joint Engn Lab Vasc Implants, Minist Educ,Bioengn Coll, Chongqing 400044, Peoples R China

[2] Chongqing Univ, Chongqing Canc Inst & Hosp, Treatment Ctr Breast Dis, Chongqing 400030, Peoples R China

[3] Chongqing Univ, Sch Microelect & Commun Engn, Chongqing Key Lab Biopercept & Intelligent Inform, Chongqing 400044, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2020年 / 163卷

基金：

中国国家自然科学基金;

关键词：

DNA methyltransferase; DNA adenine methylation methyltransferase; biological amplification techniques; CRISPR/Cpf; 1; fluorescence; DISPLACEMENT AMPLIFICATION STRATEGY; METHYLTRANSFERASE ACTIVITY; SENSITIVE DETECTION; DNA METHYLATION; BINDING; ASSAY;

D O I：

10.1016/j.bios.2020.112271

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

DNA adenine methylation methyltransferase (Dam MTase) plays an important role in gene expression and cell development, and involves in some development of tumors. There are many methods to detect Dam MTase activity, yet they have some shortcomings such as high cost, limited detection limit and complicated operation. Here, a new fast and simple multiple amplification strategy based on terminal deoxynucleotidyl transferase induced activators to unlock the collateral cleavage activities (trans-cleavage) of CRISPR/Cpf 1 (TdT-IU-CRISPR/Cpf 1) was firstly established. Based on this, a new fluorescent biosensor for Dam MTase activity detection was proposed. Importantly, the proposed biosensor does not require strict control over the temperature changes, and has ultrasensitive detection with limit of detection as low as 1.26 x 10(-3) U/mL. Moreover, the novel biosensor can not only be applied for screening the inhibitors for Dam MTase activity, but also can detect its activity in complex biological samples. The newly established multiple amplification strategy TdT-IU-CRISPR/Cpf 1 and the proposed biosensor for Dam MTase activity will be of great significance in biomedical testing and clinical diagnostics.

引用

页数：7

共 33 条

[1] ALLOSTERIC AND CATALYTIC BINDING OF S-ADENOSYLMETHIONINE TO ESCHERICHIA-COLI DNA ADENINE METHYLTRANSFERASE MONITORED BY H-3 NMR [J].