Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells

	Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells
刊名	CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
	2017
卷号	45
关键词	GENE-EXPRESSION MESSENGER-RNAS RIBOSWITCHES APTAMERS LIGANDS SYSTEM Thiamine pyrophosphate ribozyme switches Enhanced green fluorescence protein Gene expression regulation Fluorescent biosensors Mammalian cells
ISSN号	0253-3820
其他题名	Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells
英文摘要	Thiamin pyrophosphate (TPP) is a thiamine (vitamin B1) derivative and an essential cofactor in oxidative metabolism of the sugars, fatty acids and amino acids in living cells. By now, numerous TPP. dependent artificial riboswitch systems have been developed to regulate target gene expression but limited in bacteria, fungi or plant cells. Herein, the activating (switch. on) and inhibiting (switch. off) TPP. depended hammerhead ribozyme switches, which are from previous reported structures of prokaryotes screening, were investigated in mammalian cells. These ribozyme switches were inserted into the 3' UTR of the enhanced green fluorescence protein (EGFP) gene to construct the efficient ribozyme. based artificial switches through overlap extension PCR cloning. The HEK293 cells were transfected with the engineered ribozyme switches at increasing concentration of TPP. The EGFP gene. regulatory ability was analyzed with fluorescent microscope and flow cytometry. These TPP. inducible gene regulation devices showed the obvious ligand dose. dependency and excellent specificity. Two "switch. on. and one "switch. off. constructs demonstrated 3. 1 -fold or 1. 9 -fold increment and 2. 3 -fold reduction of EGFP level respectively with 150. mu mol/L TPP. The ligand. responsive ribozyme switches, by tuning the change of TPP concentration into the visual reporter genetic expression in cells, enable an efficient development of label. free, noninvasive and high. specific biosensors in living
资助项目	[National Natural Science Foundation of China] ; [Natural Science Foundation of Jiangsu Province]
语种	英语
CSCD记录号	CSCD:5918557
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/56692]
专题	中国科学院合肥物质科学研究院
推荐引用方式 GB/T 7714	. Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells[J]. CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,2017,45.
APA	(2017).Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells.CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,45.
MLA	"Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells".CHINESE JOURNAL OF ANALYTICAL CHEMISTRY 45(2017).