Abstract
This chapter describes a screening strategy to engineer synthetic riboswitches that can chemically regulate gene expression in mammalian cells. Riboswitch libraries are constructed by randomizing the key nucleotides that couple the molecular recognition function of an aptamer with the self-cleavage activity of a ribozyme. The allosteric ribozyme (aptazyme) candidates are cloned in the 3′ untranslated region (UTR) of a reporter gene mRNA. The plasmid-encoded riboswitch candidates are transfected into a mammalian cell line to screen for the desired riboswitch function. Furthermore, multiple aptazymes can be cloned into the 3′ UTR of a desired gene to obtain a logic gate response to multiple chemical signals. This screening strategy complements other methods to engineer robust mammalian riboswitches to control gene expression.
Original language | English (US) |
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Pages (from-to) | 141-148 |
Number of pages | 8 |
Journal | Methods in Molecular Biology |
Volume | 1316 |
DOIs | |
State | Published - May 16 2015 |
Externally published | Yes |
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Keywords
- Aptamer
- Aptazyme
- Gene regulation
- Riboswitch
- Ribozyme
- RNA engineering
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Aptazyme-based riboswitches and logic gates in mammalian cells. / Nomura, Yoko; Yokobayashi, Yohei.
In: Methods in Molecular Biology, Vol. 1316, 16.05.2015, p. 141-148.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Aptazyme-based riboswitches and logic gates in mammalian cells
AU - Nomura, Yoko
AU - Yokobayashi, Yohei
PY - 2015/5/16
Y1 - 2015/5/16
N2 - This chapter describes a screening strategy to engineer synthetic riboswitches that can chemically regulate gene expression in mammalian cells. Riboswitch libraries are constructed by randomizing the key nucleotides that couple the molecular recognition function of an aptamer with the self-cleavage activity of a ribozyme. The allosteric ribozyme (aptazyme) candidates are cloned in the 3′ untranslated region (UTR) of a reporter gene mRNA. The plasmid-encoded riboswitch candidates are transfected into a mammalian cell line to screen for the desired riboswitch function. Furthermore, multiple aptazymes can be cloned into the 3′ UTR of a desired gene to obtain a logic gate response to multiple chemical signals. This screening strategy complements other methods to engineer robust mammalian riboswitches to control gene expression.
AB - This chapter describes a screening strategy to engineer synthetic riboswitches that can chemically regulate gene expression in mammalian cells. Riboswitch libraries are constructed by randomizing the key nucleotides that couple the molecular recognition function of an aptamer with the self-cleavage activity of a ribozyme. The allosteric ribozyme (aptazyme) candidates are cloned in the 3′ untranslated region (UTR) of a reporter gene mRNA. The plasmid-encoded riboswitch candidates are transfected into a mammalian cell line to screen for the desired riboswitch function. Furthermore, multiple aptazymes can be cloned into the 3′ UTR of a desired gene to obtain a logic gate response to multiple chemical signals. This screening strategy complements other methods to engineer robust mammalian riboswitches to control gene expression.
KW - Aptamer
KW - Aptazyme
KW - Gene regulation
KW - Riboswitch
KW - Ribozyme
KW - RNA engineering
UR - http://www.scopus.com/inward/record.url?scp=84929314357&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84929314357&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-2730-2_12
DO - 10.1007/978-1-4939-2730-2_12
M3 - Article
C2 - 25967059
AN - SCOPUS:84929314357
VL - 1316
SP - 141
EP - 148
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -