Computational Design and Experimental Validation of Oligonucleotide Sensing Allosteric Ribozymes  

Oleh:

Penchovsky, Robert ; Breaker, Ronald R.

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 11 (Nov. 2005), page 1424-1434.
Topik: nucleotide sequence; computational design; oligonucleotide; ribozymes

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.4 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

Allosteric RNAs operate as molecular swithces that alter folding and function in response to ligand binding. A common type of natural allosteric RNAs is the riboswitch, designer RNAs with similar properties can be created by RNA engineering. We describe a computational approach for designing allosteric ribozymes triggered by binding oligonucleotides. Four universal types of RNA swithces possessing AND, OR, YES and NOT Boolean logic functions were created in modular form, which allows ligand specificity to be changed without altering the catalytic core of the ribozyme. All computationally designed allosteric ribozymes were synthesized and experimentally tested in vitro. Engineered ribozymes exhibit > 1,000 fold activation, demonstrate precise ligand specificity and function in molecular circuits in which the self-cleavage product of one RNA triggers the action of a second. This engineering approach provides a rapid and inexpensive way to create allosteric RNAs for constructing complex molecular circuits, nucleid acid detection systems and gene control elements.

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