The present invention relates to combination therapies for the treatment of a variety of disorders in mammals, including hepatic disorders and cancer. The combination of agents includes naturally-occurring (versus synthetic) oligonucleotides, particularly immunostimulatory oligodeoxynucleotides such as CpG ODNs, obtained from a natural source and one or more extracts from a Gram positive bacteria, such as Lactobacillus spp.

Disclosed are T7 RNA polymerase variants with enhanced transcriptional activity. T7 RNA polymerase variants are known which have the ability to incorporate modified ribonucleotides into growing RNA molecules. However, these variants have relatively low levels of transcriptional activity. Presented herein are mutations that increase the transcriptional activity of the variants with broad substrate range.

Methods are provided for making an RNA molecule derived from non-coding chimeric mitochondrial RNAs (ncmtRNAs), in particular antisense non-coding chimeric mitochondrial RNAs (ASncmtRNAs), compositions containing the isolated RNA molecule, methods of causing apoptosis in a cancer cell by contacting the cell with the RNA molecule, and methods of treating cancers by administering the RNA molecule to a subject in need thereof.

The invention relates to a method for production of single-stranded macronucleotides by amplifying and ligating an extended monomeric single-stranded target nucleic acid sequence (target.sub.ss) into a repetitive cluster of double-stranded target nucleic acid sequences (target.sub.ds), and subsequently cloning the construct into a vector (aptagene vector). The aptagene vector is transformed into host cells for replication of the aptagene and isolated in order to optain single-stranded target sequences (target.sub.ss). The invention also relates to single-stranded nucleic acids, produced by a method of the invention.

An insulin analogue comprises a B-chain polypeptide containing a cyclohexanylalanine substitution at position B24 and optionally containing additional amino-acid substitutions at positions A8, B28, and/or B29. A proinsulin analogue or single-chain insulin analogue containing a B domain containing a cyclohexanylalanine substitution at position B24 and optionally containing additional amino-acid substitutions at positions A8, B28, and/or B29. The analogue may be an analogue of a mammalian insulin, such as human insulin. A nucleic acid encoding such an insulin analogue is also provided. A method of lowering the blood sugar of a patient comprises administering a physiologically effective amount of the insulin analogue or a physiologically acceptable salt thereof to a patient. A method of semi-synthesis using an unprotected octapeptide by means of modification of an endogenous tryptic site by non-standard amino-acid substitutions.

The present invention relates to combination therapies for the treatment of a variety of disorders in mammals, including hepatic disorders and cancer. The combination of agents includes naturally-occurring (versus synthetic) oligonucleotides, particularly immunostimulatory oligodeoxynucleotides such as CpG ODNs, obtained from a natural source and one or more extracts from a Gram positive bacteria, such as Lactobacillus spp.

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.

A method for actively forming a three-dimensional structure, a modification mode for a nucleic acid maintaining a three-dimensional structure; and an artificial nucleic acid capable of stably binding to a target sequence regardless of mutation, by utilizing non-complementary base pairs in nucleic acid therapeutics are provided. An artificial nucleic acid for inducing a specific three-dimensional structure by hybridizing with a nucleic acid of interest that does not form a functional three-dimensional structure, a gene expression inhibiting agent and a nucleic acid detecting agent including the artificial nucleic acid as an active ingredient; and a method for producing an artificial nucleic acid are also disclosed.

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.

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. ##STR00001##