Reprogramming of cellular adhesion
Disclosed are membrane-anchored polynucleotides, and compositions comprising the membrane-anchored polynucleotides. Also disclosed are the processes for the synthesis of these compounds, compositions comprising such compounds, and the use of such compounds and compositions in research and therapeutic applications.
1. A system comprising:
(i) a first isolated cell comprising a membrane anchored polynucleotide duplex, the duplex comprising:
a first compound comprising:
a membrane anchoring region comprising a long alkyl chain of at least 12 carbon atoms inserted into a lipid bilayer of the first isolated cell; and
a polynucleotide having a membrane distal end and a membrane proximal end, wherein the polynucleotide is at least 70 nucleotides and comprises:
(1) a linker region comprising a contiguous stretch of at least 60 nucleotides, and
(2) a membrane distal adhesion region comprising at least 10 nucleotides and positioned distal to the linker region, wherein the linker region is not hybridizable to the membrane distal adhesion region;
wherein the polynucleotide is conjugated to the membrane anchoring region at the membrane proximal end; and
a second compound comprising:
a membrane anchoring region comprising a long alkyl chain of at least 12 carbon atoms inserted into the lipid bilayer of the isolated cell and conjugated to a polynucleotide that is shorter than the polynucleotide of the first compound and comprises a contiguous stretch of at least 10 nucleotides that is hybridized to the polynucleotide of the first compound to a region outside of the membrane distal adhesion region; and
(ii) a second isolated cell comprising a third compound, the third compound comprising:
a membrane anchoring region comprising a long alkyl chain of at least 12 carbon atoms inserted into a lipid bilayer of the second isolated cell;
a polynucleotide having a membrane distal end and a membrane proximal end, wherein said polynucleotide is at least 70 nucleotides and comprises:
(1) a linker region comprising a contiguous stretch of at least 60 nucleotides, and
(2) a membrane distal adhesion region comprising at least 10 nucleotides and positioned distal to the linker region, wherein the linker region is not hybridizable to the membrane distal adhesion region;
wherein the polynucleotide of the third compound is conjugated to the membrane anchoring region at the membrane proximal end of the third compound;
wherein the membrane distal adhesion region of the first compound and the membrane distal adhesion region of the third compound are hybridizable such that hybridization of the membrane distal adhesion region of the first compound and the membrane distal adhesion region of the third compound results in assembly of the first cell and the second cell with each other.
2. The system of claim 1 , wherein the polynucleotide of the first compound is a DNA polynucleotide, and the membrane proximal end is a 5′ end of the polynucleotide.
3. The system of claim 1 , wherein the membrane anchoring region of the first and second compounds of the composition comprises a monoalkyl chain comprising 16 to 18 carbon atoms.
4. The system of claim 3 , wherein the membrane anchoring region of the first and second compounds of the composition comprises a monoalkylamide.
5. The system of claim 1 , wherein the membrane anchoring region of the first and second compounds comprises a dialkylphosphoglyceride.
6. The system of claim 5 , wherein the dialkylphosphoglyceride is a C 16 dialkylphosphoglyceride.
7. The system of claim 6 , wherein the membrane distal adhesion region of the first compound comprises (CAGT) 5 or (ACTG) 5 .
8. The system of claim 7 , wherein the polynucleotide of the first compound comprises 80 to 100 nucleotides.
9. The system of claim 8 , wherein the linker region of the first compound comprises a contiguous stretch of at least 60 thymine or adenine nucleotides.
10. The system of claim 1 , wherein the linker region of the first and third compounds comprises 60 to 200 nucleotides.
11. The system of claim 1 , wherein the linker region of the first and third compounds comprises 60 to 100 nucleotides.
12. The system of claim 1 , wherein the linker region of the first and third compounds comprises 60 to 80 nucleotides.
13. The system of claim 1 , wherein the linker region of the first and third compounds comprises a contiguous stretch of 60 to 500 thymine nucleotides.
14. The system of claim 1 , wherein the linker region of the first and third compounds comprises a contiguous stretch 60 to 100 thymine nucleotides.
15. The system of claim 1 , wherein the linker region of the first and third compounds comprises a contiguous stretch of 60 to 80 thymine nucleotides.
16. The system of claim 1 , wherein the membrane anchoring region of the first and second compounds comprises a substituted aliphatic chain.
17. The system of claim 1 , wherein there are no nucleotides separating the linker region from the membrane proximal end of the polynucleotide of the first compound and the second compound.
18. The system of claim 1 , further comprising a fluorophore conjugated to the membrane distal end of the polynucleotide of the first compound.
19. The system of claim 1 , further comprising a pharmaceutical composition.
20. The system of claim 1 , wherein the membrane anchoring region of the first compound comprises dialkylphosphoglyceride comprising alkyl chains of 12-22 carbon atoms or a monoalkyl amide comprising a monoalkyl chain of 16 to 18 carbon atoms.
21. The system of claim 1 , wherein the membrane anchoring region of the second compound comprises dialkylphosphoglyceride comprising alkyl chains of 12-22 carbon atoms or a monoalkyl amide comprising a monoalkyl chain of 16 to 18 carbon atoms.
22. The system of claim 1 , wherein the contiguous stretch of at least 10 nucleotides of the second compound is hybridized to the linker region of the first compound.
23. The system of claim 1 , wherein the polynucleotide of the first compound comprises a membrane proximal adhesion region between the linker and the membrane distal adhesion region and wherein the contiguous stretch of at least 10 nucleotides of the second compound is hybridized to the membrane proximal adhesion region of the first compound.