Artificial riboswitches and other ligand-responsive gene regulators make it possible to switch protein synthesis ON or OFF with arbitrary ligand molecules. Artificial Riboswitches: Methods and Protocols focuses on the state-of-the-art methods developed in recent years for creating artificial riboswitches, therefore this volume could be regarded as a collection of recipes for the gene circuit elements in synthetic biology and metabolic engineering. Chapters cover topics such as screening or rational design methods for obtaining artificial riboswitches that function in either bacterial or eukaryotic translational systems, protocols for evaluating the activities of the resultant riboswitches, as well as protocols for construction of ligand-dependent, trans-acting gene regulators. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Artificial Riboswitches: Methods and Protocols seeks to serve not only bioengineers who aim to reprogram cell behaviors and molecular biologists who leverage these regulators for genetic studies, but to all researchers interested in this fascinating field.
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1. Computational Design of RNA Libraries for in vitro Selection of Aptamers
Yaroslav Chushak, Jennifer A. Martin, Jorge L. Chávez, Nancy Kelley-Loughnane, and Morley O. Stone
2. In vitro Selection of RNA Aptamers for a Small-Molecule Dye
Asako Murata and Shin-ichi Sato
3. Development of Photoswitchable RNA Aptamer-Ligand Complexes
Gosuke Hayashi and Kazuhiko Nakatani
4. Identification of RNA Aptamers Against Recombinant Proteins with a Hexa-Histidine Tag
Shoji Ohuchi
5. Artificial Riboswitch Selection: A FACS-Based Approach
Zohaib Ghazi, Casey C. Fowler, and Yingfu Li
6. FRET-Based Optical Assay for Selection of Artificial Riboswitches
Svetlana V. Harbaugh, Molly E. Chapleau, Yaroslav G. Chushak, Morley O. Stone, and Nancy Kelley-Loughnane
7. Portable Two-Way Riboswitches: Design and Engineering
Ye Jin and Jian-Dong Huang
8. Generation of Orthogonally Selective Bacterial Riboswitches By Targeted Mutagenesis and in vivo Screening
Helen A. Vincent, Christopher J. Robinson, Ming-Cheng Wu, Neil Dixon, and Jason Micklefield
9. Dual Genetic Selection of Synthetic Riboswitches in Escherichia coli
Yoko Nomura and Yohei Yokobayashi
10. Nucleotide Kinase-Based Selection System for Genetic Switches
Kohei Ike and Daisuke Umeno
11. Measuring Riboswitch Activity in vitro and in Artificial Cells with Purified Transcription-Translation Machinery
Laura Martini and Sheref S. Mansy
12. Rational Design of Artificial ON-Riboswitches
Atsushi Ogawa
13. Engineering Protein-Responsive mRNA Switch in Mammalian Cells
Kei Endo and Hirohide Saito
14. Guanine-Tethered Antisense Oligonucleotides as Synthetic Riboregulators
Masaki Hagihara
15. In Vitro Selection of Allosteric Ribozymes that Sense the Bacterial Second Messenger c-di-GMP
Kazuhiro Furukawa, Hongzhou Gu, and Ronald R. Breaker
16. Dual-Selection for Evolution of in vivo Functional Aptazymes as Riboswitch Parts
Jonathan A. Goler, James M. Carothers, and Jay D. Keasling
17. In vivo Screening for Aptazyme-Based Bacterial Riboswitches
Charlotte Rehm and Jörg S. Hartig
18. Engineered Riboswitch as Gene-Regulatory Platform for Reducing Antibiotic Resistance
Libing Liu and Shu Wang
19. Construction of Ligand-Responsive microRNAs that Operate Through Inhibition of Drosha Processing
Chase L. Beisel, Ryan J. Bloom, and Christina D. Smolke
20. A Three-Dimensional Design Strategy for a Protein-Responsive shRNA Switch
Shunnichi Kashida and Hirohide Saito
Le informazioni nella sezione "Su questo libro" possono far riferimento a edizioni diverse di questo titolo.
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