title: Selection of Intracellularly Functional RNA Mimics of Green Fluorescent Protein Using Fluorescence-Activated Cell Sorting.
abstract: Fluorescence-activated cell sorting (FACS) was exploited to isolate Escherichia coli cells that were highly fluorescent due to the expression of RNA aptamers that induce fluorescence of 3,5-difluoro-4-hydroxybenzylidene imidazolinone. Two different aptamers, named ZT-26 and ZT-324, were identified by this method and compared to the fluorescence-signaling properties of Spinach, a previously reported RNA aptamer. Aptamer ZT-26 exhibits significantly enhanced fluorescence over Spinach only in vitro. However, aptamer ZT-324 is 36% brighter than Spinach when expressed in E. coli. The FACS-based selection strategy presented here is attractive for deriving fluorescent RNA aptamers that function in cells as it directly selects for cells with a high level of fluorescence due to the expression of the RNA aptamer.
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Zou
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wu
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Gangyi
authors.initials: G
authors.lastname: Chen
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Juan
authors.initials: J
authors.lastname: Dong
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Cui
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, People's Republic of China. tangzhuo@cib.ac.cn.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Tang
mesh_terms: D052157:Aptamers, Nucleotide; D001593:Benzyl Compounds; D004926:Escherichia coli / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D049452:Green Fluorescent Proteins; D048288:Imidazolines; D052156:SELEX Aptamer Technique / Q000379:methods*
chemical_list: C560120:3,5-difluoro-4-hydroxybenzylidene imidazolinone; D052157:Aptamers, Nucleotide; D001593:Benzyl Compounds; D048288:Imidazolines; D049452:Green Fluorescent Proteins
keywords: Aptamers; FACS; Fluorescence; In vivo