METHODS AND COMPOSITIONS RELATED TO CHITOSAN-DERIVED NANOPARTICLE-MEDIATED CRISPR/CAS9 DELIVERY

• US 20190321477A1
• Filed: 06/27/2017
• Published: 10/24/2019
• Est. Priority Date: 06/27/2016
• Status: Active Application

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Smyth, Hugh D.C., Zhang, Hairui, Ferrati, Silvia
• Application Number:
  US16/313,255
• Publication Number:
  US 20190321477A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/36 Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
• CPC Class Codes: A61K 47/6935 the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
• CPC Class Codes: A61K 47/6939 the polymer being a polysaccharide, e.g. starch, chitosan, chitin, cellulose or pectin
• CPC Class Codes: A61K 9/5161 Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/20 involving clustered regularly interspaced short palindromic repeats [CRISPRs]
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2800/80 Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites

Abstract

Disclosed herein are nucleic acid delivery systems and methods of use.

Claims

1. A polymer carrier comprising polyethylene glycol (PEG) chemically bound to a chitosan or oligo-chitosan backbone

2. A nucleic acid delivery system comprising:

a. a CRISPR-Cas9 polynucleotide sequence, wherein the polynucleotide sequence comprises a guide sequence capable of hybridizing to a target sequence in a eukaryotic cell;

b. donor-corrected nucleic acid; and

c. a polymeric carrier system, wherein the polymeric carrier comprises polyethylene glycol (PEG) bound to a chitosan or oligo-chitosan backbone.

3. The nucleic acid delivery system of claim 2, wherein the delivery system forms nanoparticles.

4. The nucleic acid delivery system of claim 2, wherein the donor-corrected nucleic acid is siRNA.

5. The nucleic acid delivery system of claim 2, wherein the donor-corrected nucleic acid corrects an F508del CFTR mutation.

6. The nucleic acid delivery system of claim 2, wherein the delivery particles are less than 500 nm in diameter.

7. The nucleic acid delivery system of claim 2, wherein the delivery particles are less than 250 nm in diameter.

8. The nucleic acid delivery system of claim 2, wherein the delivery particles are less than 100 nm in diameter.

9. The nucleic acid delivery system of claim 2, wherein the nanoparticle formulation comprises a lipid-based nanoparticle.

10. The nucleic acid delivery system of claim 2, wherein the ratio of moles of the amine groups of chitosan to that of the phosphate of DNA (N/P ratio) is less than 1.

11. A method of modifying an organism or a non-human organism by manipulation of a target sequence in a genomic locus of interest comprising administering a composition comprising the nucleic acid delivery system of claim 2.

12. A method of treating a subject with cystic fibrosis, the method comprising administering the nucleic acid delivery system of claim 4.