Neural transfection reagents
The invention is directed to transfection reagents for the delivery of nucleic acids into neural cells, compositions including the reagents, methods of preparation of such reagents, methods of transfecting cells with such reagents, and uses thereof. In preferred embodiments the reagents comprise horseradish peroxidase and/or a polycarboxylic acid such as poly(acrylic acid) or poly(methacrylic acid).
1. A transfection reagent comprising
(a) a neural transport agent comprising an enzyme, polymer, lectin, fluorescent dye, dextran, or a combination of any two or more of these, and
(b) a nucleic acid carrier disposed within and conjugated to the neural transport agent,
wherein the nucleic acid carrier and the neural transport agent comprise complementary chemically reactive groups or high affinity ligands to conjugate the carrier and the transport agent, and
wherein the transfection reagent has a net negative surface charge under mammalian physiological conditions and an overall effective diameter of 5 to 250 nm.
2. The transfection reagent according to claim 1 , wherein the neural transport agent comprises a polycarboxylic acid.
3. The transfection reagent according to claim 1 , wherein the neural transport agent comprises an acrylic polymer.
4. The transfection reagent according to claim 3 , wherein the neural transport agent comprises poly(acrylic acid) or poly(methacrylic acid).
5. The transfection reagent according to claim 1 , wherein the nucleic acid carrier contains amines.
6. The transfection reagent according to claim 1 , wherein the transfection reagent additionally comprises a neuronal cell subtype targeting ligand.
7. The transfection reagent according to claim 1 , wherein the nucleic acid carrier is a nanoparticle whose diameter is in a range of 5 nm to 250 nm.
8. The transfection reagent according to claim 1 , wherein the nucleic acid carrier comprises a polyplex forming agent.
9. The transfection reagent according to claim 1 , wherein the nucleic acid carrier comprises a cationic polymer.
10. The transfection reagent according to claim 1 further comprising nucleic acid.
11. The transfection reagent according to claim 10 , wherein the nucleic acid is selected from RNA, DNA, siRNA, miRNA, piRNA, shRNA, antisense nucleic acid, aptamers, ribozymes, locked nucleic acid, catalytic DNA, plasmid, cosmid, artificial chromosomes, morpholinos or other synthetic nucleic acid analogs.
12. The transfection reagent according to claim 1 further comprising a passivating polymer.
13. A composition for transfection of neurons, the composition comprising a neural transport agent comprising an enzyme, polymer, lectin, fluorescent dye, dextran, or a combination of any two or more of these and a nucleic acid carrier, wherein the neural transport agent and the nucleic acid carrier are conjugated to each other via complementary chemically reactive groups or high affinity ligands, and wherein, once conjugated, the composition has a net negative surface charge under mammalian physiological conditions and an overall effective diameter of 5 to 250 nm or less.
14. The composition according to claim 13 , further comprising nucleic acid.
15. The composition according to claim 13 , wherein the neural transport agent and the nucleic acid carrier are separate but capable of forming a stable interaction when mixed together.
16. The transfection reagent according to claim 1 , wherein the nucleic acid carrier and the neural transport agent contain complementary bioorthagonal chemically reactive groups.