The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules such as chikungunya viral proteins, such as chikungunya envelope protein E1.

An electrochemical screening method for the selection of DNA aptamers against 11-deoxycortisol (11-DCL) using gold electrode for target immobilization is described. The gold electrode is used as solid matrix instead of the beads for SELEX. The selection steps (SELEX) are performed on the 11-DC modified electrode directly as the DNA library in the first round or the enriched DNA pools in the subsequent rounds were incubated on the electrode, then the unbound DNA is washed and the bound DNA is measured directly by square wave voltammetry. Then elution of the bound DNA is performed for further use.

The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules, such as drug compounds, such as levamisole and cocaine.

The present disclosure describes compositions and methods for rapid selection of both binding and functional oligonucleotides (DNA, RNA, or any natural or synthetic analog of these). In certain embodiments, provided herein are flow cells (e.g., flow cells for an Illumina sequencing instrument or a Polonator sequencing instrument) comprising within its flow chamber a plurality of immobilized aptamer clusters (e.g., from an aptamer library described herein) and, optionally, one or more target cells (e.g., cancer cells, immune cells, etc.) and/or a detectable indicator of cellular function (e.g., a fluorescent indicator of apoptosis, cell proliferation, gene or protein expression, etc.). In certain embodiments, provided herein are methods of using such an aptamer cluster-containing flow cell to identify functional aptamers from an aptamer library (e.g., in a sequencing instrument, such as an Illumina sequencing instrument).

The present invention is related to a method for generating a nucleic acid molecule capable of binding to a target molecule comprising the following steps: a) providing a reference nucleic acid molecule, wherein the reference nucleic acid molecule is capable of binding to the target molecule and wherein the reference nucleic acid molecule comprises a sequence of nucleotides, wherein the sequence of nucleotides comprises n nucleotides; b) preparing a first level derivative of the reference nucleic acid molecule, wherein the first level derivative of the reference nucleic acid molecule differs from the reference nucleic acid molecule at one nucleotide position, wherein the first level derivative is prepared by replacing the ribonucleotide at the one nucleotide position by a 2-deoxyribonucleotide in case the reference nucleic acid has a ribonucleotide at the nucleotide position and wherein the first level derivative is prepared by replacing the 2-deoxyribonucleotide at the one nucleotide position by a ribonucleotide in case the reference nucleic acid has a 2-deoxyribonucleotide at the nucleotide position and wherein the nucleotide position at which the replacement is made is the modified nucleotide position; and c) repeating step b) for each nucleotide position of the reference nucleic acid molecule, thus preparing a group of first level derivatives of the reference nucleic acid molecule, wherein the group of first level derivatives of the reference nucleic acid molecule consists of n first level derivatives, wherein each of the first level derivatives of the reference nucleic acid molecule differs from the reference nucleic acid molecule by a single nucleotide replacement and wherein each of the first level derivatives of the reference nucleic acid molecule has a single modified nucleotide position which is different from the single modified nucleotide of all of the single modified nucleotide positions of the other first level derivatives of the group of first level derivatives of the reference nucleic acid molecule.

The present disclosure provides methods, kits and compositions for identifying nucleic acid agents having a desired property, e.g., a property of specifically binding to a target (such as a protein target) with high affinity. More specifically, the present disclosure provides methods, kits and compositions for identifying candidate nucleic acid agents with both high specificity and affinity for a target.

Efficient sequence specific gene silencing is possible through the use of siRNA technology. Be selecting particular siRNAs by rational design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes. Methods compositions, and kits generated through rational design of siRNAs are disclosed, including those directed to the nucleotide sequences for F12.

Methods and compositions are provided for specific aptamers and aptamer pools that bind biomarkers of interest such as microvesicle surface antigens or functional fragments of microvesicle surface antigens. In various embodiments, aptamers of the invention are used in diagnostic, prognostic, or theranostic processes to screen a biological sample for the presence or levels of biomarkers such as microvesicles that are determined to provide a diagnostic, prognostic, or theranostic readout. The diagnosis, prognosis, or theranosis may be related to cancer or other diseases and disorders. The invention also provides methods and composition to facilitate aptamer library screening and aptamer detection methods.

Provided herein is technology relating to the mutagenesis of nucleic acids, e.g., for directed evolution, and particularly, but not exclusively, to methods, compositions, and kits for producing nucleic acids and/or proteins comprising mutations and substitutions within specific target sequences.

Methods of generating novel guide nucleic acids comprising a template-conserved target complementary region to a template and template-randomized region, novel guide nucleic acids generated by the methods, mixtures and complexes comprising the novel guide nucleic acids are disclosed.