Distinguishing PCA3 messenger RNA species in benign and malignant prostate tissues
View Patent ↗This invention concerns the discovery of two distinct PCA3 mRNA sequences. One of these sequences corresponds to a short PCA3 mRNA molecule whereas the other PCA3 RNA molecule is longer as it comprises an additional sequence between exon 3 and exon 4 a . The short RNA is associated with prostate cancer whereas the long RNA sequence is associated with a non-malignant state of the prostate. Based on the differential expression levels of these two PCA3 RNA sequences, protocols for the diagnosis of prostate disease are provided. The invention also relates to therapeutic approaches to prostate cancer.
1. A cDNA molecule encoding a differentially expressed prostate cancer antigen 3 (PCA3) mRNA containing an additional sequence between exon 3and exon 4a, thereby giving rise to a long PCA3 mRNA, wherein the additional sequence is at least 90% identical to the sequence of positions 27-254 of SEQ ID NO: 1 or the complement thereof.
2. The cDNA molecule of claim 1 , wherein said additional sequence interrupts the open reading frame of a PCA3 protein, thereby yielding the truncated PCA3 protein.
3. The cDNA molecule according to claim 1 , comprising a polynucleotide sequence at least 90% identical to the nucleotide sequence as set forth in SEQ ID NO:1 or the complement thereof.
4. The cDNA molecule according to claim 1 , wherein the molecule comprises the nucleotide sequence encoding PCA3 as set forth in SEQ ID NO:1.
5. A recombinant nucleic acid molecule comprising 5′ to 3′, a promoter effective to initiate transcription in a host cell and the cDNA molecule according to claim 1 .
6. A cell that contains the recombinant nucleic acid molecule according to claim 5 .
7. A non-human organism that contains the recombinant nucleic acid molecule according to claim 5 .
8. The cDNA molecule according to claim 1 , wherein the additional sequence is at least 95% identical to the sequence of positions 27-254 of SEQ ID NO: 1 or the complement thereof.
9. The cDNA molecule according to claim 1 , wherein the additional sequence is at least 98% identical to the sequence of positions 27-254 of SEQ ID NO: 1 or the complement thereof.
10. The cDNA molecule according to claim 1 , wherein the additional sequence comprises the sequence of positions 27-254 of SEQ ID NO: 1 or the complement thereof.
11. A composition comprising the cDNA of claim 1 and at least one oligonucleotide comprising a sequence complementary to at least 12 consecutive nucleotides of the cDNA of claim 9 .
12. The composition of claim 11 , wherein the oligonucleotide is a primer.
13. The composition of claim 11 , wherein the composition comprises a pair of primers for amplifying an amplicon from the cDNA.
14. The composition of claim 11 , wherein the oligonucleotide is a labeled probe.
15. The composition of claim 11 , wherein the oligonucleotide is a fluorescently labeled probe.
16. A composition comprising the cDNA of claim 1 and at least one DNA polymerase.
17. The composition of claim 16 , wherein the DNA polymerase is a reverse transcriptase or a thermostable DNA polymerase.
18. The composition of claim 16 , further comprising at least one primer.