Property effecting and/or property exhibiting compositions for therapeutic and diagnostic uses
The present invention provides an array of compositions useful for effecting and/or exhibiting changes in biological functioning and processing within cells and in biological systems containing such cells. In effect, these compositions combine chemical modifications and/or ligand additions with biological functions. The chemical modifications and/or ligand additions provide additional characteristics to the compositions without interfering substantially with their biological function. Such additional characteristics include nuclease resistance, targeting specific cells or specific cell receptors localizing to specific sites within cells and augmenting interactions between the compositions and target cells of interest as well as decreasing such interactions when desired. Also provided by the present invention are processes and kits.
1. A chemically modified nucleic acid construct, said construct comprising
(i) at least one nucleic acid strand consisting of unmodified nucleotides; and
(ii) one or more nucleic acid strands comprising one or more modified nucleotides or nucleotide analogs, wherein
at least one modified nucleotide or nucleotide analog comprises a fusogenic peptide,
at least one modified nucleotide or nucleotide analog comprises a ligand to a cell receptor, and
at least one modified nucleotide or nucleotide analog comprises a non-nucleic acid entity that confers nuclear localization,
and wherein said at least one nucleic acid strand consisting of unmodified nucleotides is a template for the synthesis of a nucleic acid product comprising RNA and having biological activity.
2. The construct of claim 1 , wherein at least a portion of said construct is linear, and at least a portion of said construct is circular.
3. The construct of claim 1 , wherein at least a portion of said construct is a first polynucleotide sequence comprising a gapped circle.
4. The construct of claim 3 , wherein said gapped portion of said gapped circle is hybridized to a second polynucleotide sequence comprising the fusogenic peptide.
5. The construct of claim 1 , having at least one 3′ terminus, said terminus comprising a polynucleotide tail.
6. The construct of claim 5 , wherein said polynucleotide tail is hybridized to a complementary polynucleotide sequence.
7. The construct of claim 1 , wherein said construct comprises DNA.
8. The construct of claim 1 , wherein at least one of said modified nucleotide or nucleotide analog has been modified at least on the sugar moiety.
9. The construct of claim 6 , wherein said ligand to a cell receptor is attached to the complementary polynucleotide sequence.
10. The construct of claim 1 , wherein said ligand to a cell receptor is a hormone, lectin, protein, polysaccharide, polysaccharide, asialoorosomucoid, transferrin or a tri-lactyl lysyl lysine.
11. A chemically modified nucleic acid construct, said construct comprising a modified nucleotide or a nucleotide analog, which construct when present in a cell is used as a template for the synthesis of a nucleic acid product comprising RNA and having biological activity, wherein
said modified nucleotide or nucleotide analog comprises at least one non-nucleic acid entity conferring cell targeting,
wherein said construct further comprises at least three strands, wherein
the first strand is a template strand comprising a promoter,
the second strand
(a) has at least one terminus, said terminus comprising a polynucleotide tail and
(b) comprises two segments, wherein the first segment is complementary and hybridizes to a portion of the first strand and the second segment lacks said complementarity and comprises said polynucleotide tail, and
the third strand comprises (i) said modified nucleotide or nucleotide analog comprising said non-nucleic acid entity conferring cell targeting, and (ii) a polynucleotide sequence hybridized to a complementary sequence in said polynucleotide tail.
12. The construct of claim 1 , wherein said construct further comprises a modified nucleotide or nucleotide analog comprising a non-nucleic acid entity comprising a nuclear localization signal.
13. The construct of claim 1 , wherein at least one of said modified nucleotide or nucleotide analog has been modified at least on the base moiety.
14. The construct of claim 1 , wherein at least one of said modified nucleotide or nucleotide analog has been modified at least on the phosphate moiety.
15. The construct of claim 11 , further comprising a modified nucleotide or nucleotide analog that comprises a non-nucleic acid entity that confers nuclear localization.
16. The construct of claim 11 , further comprising a modified nucleotide or nucleotide analog that comprises a non-nucleic acid entity that confers nuclear localization.
17. The construct of claim 1 , wherein said construct comprises RNA.
18. The construct of claim 11 , wherein the first strand comprises a gapped circle having a double stranded portion and a single stranded portion, wherein said complementary portion of the second strand is hybridized to the single stranded portion.