Trans-splicing rna (tsrna)
AssigneeNational University of Singapore
InventorsPei She Loh, Volker Patzel
Life of this patent
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Abstract
Trans-splicing RNA (tsRNA) The invention concerns a trans-splicing RNA (tsRNA) molecule comprising at least one or multiple unstructured target binding domains complementary to at least one or multiple precursor messenger RNA (pre-mRNA) targets and adapted to prevent off-target trans-splicing by the inclusion of a safety domain; a cell or vector or therapeutic or composition or pharmaceutical composition comprising said tsRNA; and a method for killing cells or treating a disease or for imaging or for a cosmetic application using said tsRNA. The invention has use in the medical, cosmetic, and veterinary field.
Claims48
A trans-splicing RNA (tsRNA) molecule comprising:
- (a) at least one binding domain specific for at least a part of a gene that associates with or is a biomarker for a cell or a disease to be treated;
- and
- (b) nucleic acids encoding at least one or more expressible
- (c) suicide protein or a protein that is a component of a suicide system;
- or
- (d) fluorescent protein, luciferase or other reporter protein;
- or
- (e) therapeutic protein;
- and
- (f) at least one splice signal;
- and
- (g) at least one safety domain specific for the splice signal or splice site within the trans-splicing RNA.
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A dumbbell-shaped DNA expression vector comprising:
- a) one or more linear or hairpin-shaped transcription cassettes each comprising a nucleotide sequence encoding a nucleic acid molecule to be expressed;
- b) two single-stranded DNA loops;
- c) operably linked to said transcription cassette a minimal transcription promoter nucleotide sequence and a transcriptional terminator;
- d) a nucleotide sequence comprising a DNA sequence that functions as nuclear targeting sequence (NTS);
- e) a nucleotide sequence comprising a spliceable intron;
- and
- f) at least one residue or helper function for targeted delivery covalently or non-covalently linked to at least one of the loops;
- optionally
- wherein said residue or helper function is selected from the group comprising:
- a carbohydrate, a (GalNAc)3 residue, a nucleic acid, a RNA or DNA or peptide aptamer, a CD137 or a PSMA binding RNA aptamer, a protein, a peptide, a cell penetrating peptide, an antibody, or a CD137-binding antibody;
- and/or
- wherein said helper function is non-covalently attached to one or both of the dumbbells loops via complementary antisense oligonucleotides (RNA or DNA) to which the helper function is covalently attached at the 5′ or 3′ end;
- and/or
- wherein said helper function is covalently attached to one or both of the dumbbells loops;
- and/or
- wherein said NTS comprises a binding site for a transcription factor;
- and/or
- wherein said NTS is the SV40 enhancer sequence, the minimal SV40 enhancer sequence, the smooth muscle cell y-actin (SMGA) promoter or the oriP of the Epstein-Barr virus (EBV).
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Dependencies are inferred from the claim text.