The present disclosure relates to methods, compositions and kits for synthesizing moderate length RNAs (mlRNAs, including gRNAs) by splint-mediated ligation of RNA fragments. The synthesis of moderate length RNAs can be followed by DNase treatment. In some embodiments, splint DNA oligonucleotides that are no longer than 32 nucleotides are used.

Provided herein, inter alia, are double stranded oligonucleotide molecules and methods of making the molecules. The double stranded oligonucleotide molecules include a first oligonucleotide strand comprising a first nucleic acid sequence bound to a second nucleic acid sequence through a first spacer, wherein said second nucleic acid sequence is bound to a third nucleic acid sequence through a second spacer and a second oligonucleotide strand comprising a fourth nucleic acid sequence bound to a fifth nucleic acid sequence through a third spacer, wherein said fifth nucleic acid sequence is bound to a sixth nucleic acid sequence through a fourth spacer, wherein the second nucleic acid sequence and the fifth nucleic acid sequence are hybridized to form a double stranded nucleic acid core of said double stranded oligonucleotide.

An object of the present invention is to provide a method for stabilizing a nucleic acid oligomer solution having a phosphorothioate bond, and a method for producing a purified nucleic acid oligomer. The present invention provides a method for stabilizing a nucleic acid oligomer, wherein an atmosphere in contact with an eluted fraction obtained by subjecting a crude nucleic acid oligomer containing a nucleic acid oligomer represented by Formula (1) (wherein symbols are as described in the specification) to a reverse-phase column chromatography treatment is set to an inert gas atmosphere having an oxygen concentration of 10% or less, and a method for producing a purified nucleic acid oligomer from the eluted fraction.

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Compositions and methods are provided for forming a single RNA polynucleotide from a plurality of DNA oligonucleotides in a single reaction chamber using combined reagents in a single step reaction. DNA polymerase, RNA polymerase and single stranded (ss) DNA oligonucleotides are combined where each DNA oligonucleotide has one or more sequence modules, wherein one sequence module in the first ss DNA oligonucleotide is complementary to a sequence module at the 3′ end of the second ss DNA oligonucleotide; and wherein a second module on the first ss DNA oligonucleotide is an RNA polymerase promoter sequence; and forming a single RNA polynucleotide, excluding the RNA promoter sequence, derived from the first and second DNA oligonucleotides

The present invention relates to new aptamer molecules for use in the treatment and/or diagnosis of autoimmune diseases associated with autoantibodies against G-protein coupled receptors, a pharmaceutical composition comprising such aptamer molecules, an apheresis column comprising such aptamer molecules and a method for the determination of nucleotide sequences for use as sequences of aptamer molecules.

A method for producing lipid particles, including: injecting molten lipids directly into a liquid at a temperature lower than a melting point of the lipids through a liquid supply port of a two-fluid nozzle while injecting a gas directly into the liquid through a gas supply port of the two-fluid nozzle, so that the molten lipids are dispersed and atomized into particles in the liquid due to the gas and the particles are solidified to form lipid particles. The lipids have a water solubility of 10 g/L or less at 25° C. and are solid at 25° C. The two-fluid nozzle is heated to a temperature at least 10° C. higher than the melting point of the lipids. A ratio D50/Nd of a volume median diameter D50 of the lipid particles to an orifice diameter Nd of the liquid supply port of the two-fluid nozzle is 0.0017 or more and 0.17 or less.

Compositions and methods are provided for forming a single RNA polynucleotide from a plurality of DNA oligonucleotides in a single reaction chamber using combined reagents in a single step reaction. DNA polymerase, RNA polymerase and single stranded (ss) DNA oligonucleotides are combined where each DNA oligonucleotide has one or more sequence modules, wherein one sequence module in the first ss DNA oligonucleotide is complementary to a sequence module at the 3′ end of the second ss DNA oligonucleotide; and wherein a second module on the first ss DNA oligonucleotide is an RNA polymerase promoter sequence; and forming a single RNA polynucleotide, excluding the RNA promoter sequence, derived from the first and second DNA oligonucleotides.

The invention discloses a method for constructing functional exosomes capable of efficiently loading specific miRNA. In order to enable the exosome to carry miRNA with specific regulation function more efficiently so as to play a role in targeted regulation more accurately and efficiently, MS2 phage capsid protein is utilized to edit and construct a capture element of a specific miRNA molecule, and placenta mesenchymal stem cells are reprogrammed to enable the secreted exosome to efficiently load a target miRNA molecule, so that the target miRNA molecule is delivered to tissue cells to play a role in effective regulation, and therefore a new strategy is provided for realizing specific precise treatment in the future.

A synthesis method of a targeted drug nCoVshRNA.2ACE2 of a COVID-19 virus, which includes the following steps: designing a consensus RNAi sequence siRNA of the COVID-19 virus and a variant strain thereof; synthesizing two complementary siRNAs into a small hairpin-shaped shRNA with a loop, and synthesizing ACE2 or a cell penetrating peptide ACE2 with a receptor-binding domain (RBD) as a ligand; and ligating the ACE2 to a sense strand and an antisense strand of the shRNA separately to synthesize the nCoVshRNA.2ACE2 including a shRNA region and an ACE2 region. The bivalent ACE2 functions to neutralize the RBD and deliver the shRNA in a targeted manner; an “shRNA-ACE2-RBD-virus” complex bridged by the ACE2 allows the shRNA to enter target cells with virus infection, thereby avoiding a side effect of non-specific delivery of the shRNA to uninfected cells, as well as resisting the variant strain and neutralizing the virus with the ACE2.

This invention provides a nucleic acid that enables evaluation of various endocrine disrupting actions of a very small amount of endocrine disrupting chemicals with high sensitivity. Such nucleic acid comprises a total of 20 to 60 nucleotides comprising the nucleotide sequence shown in SEQ ID NO: 1 and shows excellent responsiveness to various endocrine disrupting chemicals.