Concatemeric RNA molecules, compositions, and methods and uses thereof

A concatemeric RNA molecule, comprising at least a first and a second repeat units, each of the first and the second repeat units including:

• a first segment having a first nucleotide sequence;
• a second segment having a second nucleotide sequence, complementary to the first nucleotide sequence;
• and
• a connecting segment covalently attached to the second segment,
• wherein at least one of the first or the second repeat units further includes a loop segment having a loop nucleotide sequence, the loop segment being covalently attached to the first and the second segments,
• wherein the loop nucleotide sequence includes at least one enzyme-specific nucleotide sequence;
• said enzyme-specific nucleotide sequence is excluded from the connecting segment.

A method for generating a concatemeric RNA molecule comprising:

• (a) providing a DNA template;
• (b) transcribing said DNA template by rolling circle transcription (RCT) or rolling circle amplification (RCA) under appropriate conditions to yield a plurality of concatemeric RNA molecules;
• wherein the RNA molecule comprises at least a first and a second repeat units,
• each of the first and the second repeat units including:
• a first segment having a first nucleotide sequence;
• a second segment having a second nucleotide sequence, complementary to the first nucleotide sequence;
• a connecting segment covalently attached to the second segment, wherein the connecting segment of the first repeat unit is covalently attached to the first segment of the second repeat unit;
• and
• a loop segment having a loop nucleotide sequence, the loop segment being covalently attached to the first and the second segments, wherein the loop nucleotide sequence includes at least one enzyme-specific nucleotide sequence;
• said enzyme-specific nucleotide sequence is excluded from the connecting segment and
• (c) isolating the plurality of concatemeric RNA molecules.