细胞功能测定

SELEX (systematic evolution of ligands by exponential enrichment) is a process for isolating DNA or RNA sequences with high affinity and selectivity for molecular targets from random sequence libraries. These sequences are commonly referred to as aptamers. The process typically re ...

Oligonucleotides complementary to RNA sequences interact poorly with folded target regions. In vitro selection of oligonucleotides carried out against RNA structures have led to aptamers that frequently differ from antisense sequences, but rather take advantage of non-doub ...

Many signalling proteins involved in diverse functions such as cell growth and differentiation can act as oncogenes and cause cellular transformation. These molecules represent attractive targets for cancer diagnosis or therapy and therefore are subject to intensive investi ...

RNA sequences fold in a hierarchical manner to form complex structures. This folding pathway proceeds first with formation of secondary structure elements followed by the compilation of tertiary contacts. Although bioinformatics-based tools are commonly used to predict secon ...

Fluorescence correlation spectroscopy (Bacia and Schwille (2007) Nat. Protoc. 2, 2842–2856) reveals molecular mobilities, enabling to identify molecular interactions based on a change of diffusion times (Rigler and Elson, (2001) Fluorescence Correlation Spectroscopy: The ...

Structural biology plays a central role in gaining a full understanding of the myriad roles of RNA in biology. In recent years, innovative approaches in RNA purification and crystallographic methods have lead to the visualization of an increasing number of unique structures, providing n ...

Cell physiology depends on a fine-tuned network of protein–protein interactions, and misguided interactions are often associated with various diseases. Consequently, peptides, which are able to specifically interfere with such adventitious interactions, are of high inter ...

The peptide aptamer approach employs high-throughput selection to identify members of a randomized peptide library displayed from a scaffold protein by virtue of their interaction with a target molecule. Extending this approach, we have developed a peptide aptamer scaffold prote ...

In vitro genetic approaches are powerful solutions to the protein design problem. mRNA display is an in vitro selection technique enabling the design of peptide and protein ligands ranging from one to over 100 amino acids. Libraries containing more than 10 trillion unique sequences can be syn ...

Peptide aptamers are small recombinant proteins typically inserted into a supportive protein scaffold. These short peptide domains can bind to their target proteins with high specificity and affinity, often resulting in an altered target protein. We describe high-throughput pro ...

Peptide aptamers have primarily been used as research tools to manipulate protein function and study regulatory networks. However, they also find multiple applications in therapeutic research, from target identification and validation to drug discovery. Because of their unbia ...

Microbodies™ are novel pharmacophoric entities which are derived from naturally occurring cystine-knot microproteins. They provide extremely stable scaffolds that can be engineered to high-affinity binding proteins. A peptide-grafting approach yielded specific liga ...

Riboswitches are RNA elements capable of modulating gene expression through interaction with cellular metabolites. One member of the riboswitch family, the glmS riboswitch, is unique among riboswitches in that it modulates gene expression by undergoing self-cleavage in the pres ...

RNA sequences fold upon themselves to form complex structures. Functional analysis of most biological RNAs requires knowledge of secondary structure arrangements and tertiary base interactions. Therefore, rapid and comprehensive methods for assessing RNA structure are hi ...

A detailed understanding of cellular mechanisms requires knowledge of structure and dynamics of the involved biomacromolecules at atomic resolution. NMR spectroscopy uniquely allows determination of static and dynamic processes at atomic level, including structured sta ...

Riboswitches are functional RNA molecules that control gene expression through conformational changes in response to small-molecule ligand binding. In addition, riboswitch 3D structure, like that of other RNA molecules, is dependent on cation–RNA interactions as the RNA backbo ...

Procedures that were critical for crystallization of the glmS ribozyme-riboswitch RNA domain from the thermophilic Gram-positive bacterium Thermoanaerobacter tengcongensis are described. Experimental design based on screening multiple variant RNA sequences and te ...

The versatility of RNA as a regulatory molecule has become increasingly apparent in recent years. RNA elements within a transcript can sense a large variety of physiological signals, resulting in modulation of expression of the gene(s) encoded on that transcript by effects on transcript sy ...

Enzymatic probing and lead(II)-induced cleavages have been developed to study the secondary structure of RNA molecules either free or engaged in complex with different ligands. Using a combination of probes with different specificities (unpaired vs. paired regions), it is possible to ...

Toeprinting was developed to study the formation of ribosomal initiation complexes in bacteria. This approach, based on the inhibition of reverse transcriptase elongation, was used to monitor the effect of ribosomal components and translational factors on the formation of the acti ...