The present invention is concerned with a method of producing a target RNA using a circular DNA, wherein said method does not comprise a step of linearizing said circular DNA. The present invention further relates to a circular DNA comprising an RNA polymerase promoter sequence, followed by a sequence encoding a target RNA, followed by a sequence encoding a self-cleaving ribozyme, followed by an RNA polymerase terminator sequence element, wherein the latter element may comprise several RNA polymerase terminator sequences. Multimeric DNA obtained by rolling circle amplification of said circular DNA is also within the scope of the present invention.

The present invention is concerned with a method of producing a target RNA using a circular DNA, wherein said method does not comprise a step of linearizing said circular DNA. The present invention further relates to a circular DNA comprising an RNA polymerase promoter sequence, followed by a sequence encoding a target RNA, followed by a sequence encoding a self-cleaving ribozyme, followed by an RNA polymerase terminator sequence element, wherein the latter element may comprise several RNA polymerase terminator sequences. Multimeric DNA obtained by rolling circle amplification of said circular DNA is also within the scope of the present invention.

Nucleobase amino acid polymer compositions and methods for synthesizing nucleobase amino acid polymers and their conversion to nucleic acids are provided. The nucleobase amino acid polymer compositions disclosed herein comprise nucleobase amino acid monomers that comprise a linker and a nucleobase. The methods disclosed herein comprise synthesizing a nucleic acid polymer by providing a mRNA template, at least one ribosome, at least one nucleobase amino acid tRNA and at least one non-nucleobase amino acid tRNA; and adding a polymerase and at least one primer to the nucleobase amino acid polymer.

Nucleobase amino acid polymer compositions and methods for synthesizing nucleobase amino acid polymers and their conversion to nucleic acids are provided. The nucleobase amino acid polymer compositions disclosed herein comprise nucleobase amino acid monomers that comprise a linker and a nucleobase. The methods disclosed herein comprise synthesizing a nucleic acid polymer by providing a mRNA template, at least one ribosome, at least one nucleobase amino acid tRNA and at least one non-nucleobase amino acid tRNA; and adding a polymerase and at least one primer to the nucleobase amino acid polymer.

The present invention relates to the field of RNA analysis. In particular, the invention concerns the use of a catalytic nucleic acid molecule for the analysis of an RNA molecule and/or of a population of RNA molecules. In one aspect, the invention concerns methods for analyzing RNA molecules having at least one cleavage site for at least one catalytic nucleic acid molecule. In particular, the invention concerns a method for determining a physical property of an RNA molecule by analyzing a 5 terminal fragment, a 3 terminal fragment and/or at least one optional central RNA fragment obtained by cleavage of the RNA molecule by at least one catalytic nucleic acid molecule. Moreover, the present invention provides novel uses of a catalytic nucleic acid molecule for analyzing RNA molecules. In particular, the invention relates to the use of a catalytic nucleic acid molecule in a method for analyzing RNA molecules, wherein the resulting 5 terminal RNA fragment, the 3 terminal RNA fragment and/or the at least one optional central RNA fragment are analyzed.

The present invention relates to the field of RNA analysis. In particular, the invention concerns the use of a catalytic nucleic acid molecule for the analysis of an RNA molecule and/or of a population of RNA molecules. In one aspect, the invention concerns methods for analyzing RNA molecules having at least one cleavage site for at least one catalytic nucleic acid molecule. In particular, the invention concerns a method for determining a physical property of an RNA molecule by analyzing a 5 terminal fragment, a 3 terminal fragment and/or at least one optional central RNA fragment obtained by cleavage of the RNA molecule by at least one catalytic nucleic acid molecule. Moreover, the present invention provides novel uses of a catalytic nucleic acid molecule for analyzing RNA molecules. In particular, the invention relates to the use of a catalytic nucleic acid molecule in a method for analyzing RNA molecules, wherein the resulting 5 terminal RNA fragment, the 3 terminal RNA fragment and/or the at least one optional central RNA fragment are analyzed.

This disclosure relates to particles conjugated to therapeutic nucleic acids. In certain embodiments, the nucleic acid comprises a sequence that catalytically cleaves RNA, e.g., DNAzyme or RNAzyme. In certain embodiments, the particles contain nucleic acids with both DNAzyme and/or RNAzyme and siRNA sequences. The cleaving nucleic acids optionally comprise a sequence functioning to hybridize to a target of interest and/or the particles are further conjugated to a targeting moiety. In certain embodiments, conjugated particles are used in the treatment or prevention of cancer or viral infections or bacterial infections. In certain embodiments, conjugated particles are used in detecting metal ions and other small molecule analyte.

This disclosure relates to particles conjugated to therapeutic nucleic acids. In certain embodiments, the nucleic acid comprises a sequence that catalytically cleaves RNA, e.g., DNAzyme or RNAzyme. In certain embodiments, the particles contain nucleic acids with both DNAzyme and/or RNAzyme and siRNA sequences. The cleaving nucleic acids optionally comprise a sequence functioning to hybridize to a target of interest and/or the particles are further conjugated to a targeting moiety. In certain embodiments, conjugated particles are used in the treatment or prevention of cancer or viral infections or bacterial infections. In certain embodiments, conjugated particles are used in detecting metal ions and other small molecule analyte.

The embodiments disclosed herein utilized RNA targeting effectors to provide a robust CRISPR-based diagnostic with attomolar sensitivity. Embodiments disclosed herein can detect both DNA and RNA with comparable levels of sensitivity and can differentiate targets from non-targets based on single base pair differences. Moreover, the embodiments disclosed herein can be prepared in freeze-dried format for convenient distribution and point-of-care (POC) applications. Such embodiments are useful in multiple scenarios in human health including, for example, viral detection, bacterial strain typing, sensitive genotyping, and detection of disease-associated cell free DNA.

The embodiments disclosed herein utilized RNA targeting effectors to provide a robust CRISPR-based diagnostic with attomolar sensitivity. Embodiments disclosed herein can detect both DNA and RNA with comparable levels of sensitivity and can differentiate targets from non-targets based on single base pair differences. Moreover, the embodiments disclosed herein can be prepared in freeze-dried format for convenient distribution and point-of-care (POC) applications. Such embodiments are useful in multiple scenarios in human health including, for example, viral detection, bacterial strain typing, sensitive genotyping, and detection of disease-associated cell free DNA.