Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\26258292
.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 Nat+Methods
2015 ; 12
(10
): 989-94
Nephropedia Template TP
Nat Methods
2015[Oct]; 12
(10
): 989-94
PMID26258292
show ga
Methods for rapidly assessing sequence-structure-function landscapes and
developing conditional gene-regulatory devices are critical to our ability to
manipulate and interface with biology. We describe a framework for engineering
RNA devices from preexisting aptamers that exhibit ligand-responsive ribozyme
tertiary interactions. Our methodology utilizes cell sorting, high-throughput
sequencing and statistical data analyses to enable parallel measurements of the
activities of hundreds of thousands of sequences from RNA device libraries in the
absence and presence of ligands. Our tertiary-interaction RNA devices performed
better in terms of gene silencing, activation ratio and ligand sensitivity than
optimized RNA devices that rely on secondary-structure changes. We applied our
method to build biosensors for diverse ligands and determine consensus sequences
that enable ligand-responsive tertiary interactions. These methods advance our
ability to develop broadly applicable genetic tools and to elucidate the
underlying sequence-structure-function relationships that empower rational design
of complex biomolecules.
free
free
free
