Nucleic acid products and methods of administration thereof

Abstract

The present invention relates in part to nucleic acids, including nucleic acids encoding proteins, therapeutics and cosmetics comprising nucleic acids, methods for delivering nucleic acids to cells, tissues, organs, and patients, methods for inducing cells to express proteins using nucleic acids, methods, kits and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, therapeutics, and cosmetics produced using these methods, kits, and devices.

1 .- 64 . (canceled) 
     
     
         65 . A method for treating cancer comprising:
 (a) providing an isolated cell;   (b) transfecting the cell with a synthetic RNA encoding a catalytic enzyme capable of converting a prodrug into an active drug, to result in the cell expressing the catalytic enzyme;   (c) administering the cell to a cancer patient; and   (d) administering the prodrug to the cancer patient, to result in conversion of the prodrug into an active drug.   
     
     
         66 . The method of  claim 65 , wherein the prodrug is selected from the group consisting of 5-Ethynyl-2(1H)-pyrimidinone, IPdR, 5-FP, d-alanine, SeCys conjugates, Menadione, Mitomycin C, Tirapazamine, EO9, Streptonigrin, CB 1954, Diaziquone, Ipomeanol, Ftorafur (tegafur), 5′-DFUR, Dacarbazine, Trofosfamide, Ifosfamide, Cyclophosphamide, AQ4N, 2,4-Dihydroxyphenylalanine, 4-S-CAP, GHB, Substituted phenols, Phenyl mustards, Urea mustards, TER286, S-CPHC-ethylsulfoxide, PTA, CPT-11, Paclitaxel-2-ethylcarbonate, Capecitabine, Tertiary amidomethyl esters, Amifostine, 3-AP phosphate, BHAMG, Epirubicin-glucuronide, HMR 1826, DNR-GA3, DOX-GA3, Paclitaxel glucuronide, 5-FU glucuronide, 5-FC, PC, GC, SeCys conjugate, 4-Ipomeanol, Fludarabine, MeP-dR, Ganciclovir, Etoposide phosphate, Mitomycin C phosphate, POMP, N-(4-phosphonooxy)-phenylacetyl)doxorubicin, Glucuronidated Nornitrogen mustard, Glucuronidated 9-amino-camptothecin, Glucuronide mustard, Methotrexate-amino acids, CMDA, DPO, N-(phenyacetyl) doxorubicin, MelPO, NHPAP, N-(phenylacetyl) doxorubicin, N-(phenylacetyl) melphalan, LY 266070, C-DOX, PRODOX, CM, CCM, Cephalosporin-DACCP, PROTAX, Cephalosporin mitomycin C, C-Mel, 5-Fluorocytosine, Selenomethionine, and Trifluoromethionine. 
     
     
         67 . The method of  claim 66 , wherein the catalytic enzyme is selected from the group consisting of an aldehyde oxidase, an amino acid oxidase, a cytochrome P450 reductase, a DT-diaphorase, a cytochrome P450, a tyrosinase, a glutathione an S-transferase, a carboxylesterase, an alkaline phosphatase, a β-glucuronidase, a cysteine conjugate β-lyase, a nitroreductase, a purine-nucleoside phosphorylase, a thymidine phosphorylase, a thymidine kinase, a carboxypeptidase, a penicillin amidase, a β-Lactamase, a cytosine deaminase, and a methionine γ-lyase. 
     
     
         68 . The method of  claim 65 , wherein the catalytic enzyme is capable of catalyzing conversion of the prodrug selected from the group consisting of 5-FP, tegafur, 5′-DFUR, 5-FU glucuronide, and 5-FC, into the active drug 5-FU, and wherein the catalytic enzyme is selected from the group consisting of an aldehyde oxidase, a cytochrome P450, a thymidine phosphorylase, a β-glucuronidase, and a cytosine deaminase. 
     
     
         69 . The method of  claim 68 , wherein the catalytic enzyme comprises the cytosine deaminase, and the cytosine deaminase is capable of catalyzing conversion of 5-FC into 5-FU. 
     
     
         70 . The method of  claim 68 , wherein the catalytic enzyme comprises the aldehyde oxidase that is capable of catalyzing conversion of 5-FC into 5-FU, or the catalytic enzyme comprises the cytochrome P450 that is capable of catalyzing conversion of tegafur into 5-FU, or the catalytic enzyme comprises a carboxylesterase that is capable of catalyzing conversion of a capecitabine into 5-FU, or the catalytic enzyme comprises the β-glucuronidase that is capable of catalyzing conversion of 5-FU glucoronide into 5-FU. 
     
     
         71 . The method of  claim 67 , wherein the catalytic enzyme is capable of catalyzing conversion of the prodrug selected from the group consisting of N-(4-phosphono-oxy)-phenylacetyl) doxorubicin, HMR 1826, DOX-GA3, DPO, N-(phenyacetyl) doxorubicin, C-DOX, and PRODOX into the active drug 5-FU, and wherein the catalytic enzyme is selected from the group consisting of the alkaline phosphatase, the β-glucuronidase, the penicillin amidase, and the β-Lactamase. 
     
     
         72 . The method of  claim 65 , wherein the synthetic RNA comprises a combination of non-canonical nucleotides that avoid substantial cellular toxicity, optionally selected from 5-hydroxycytidine, 5-methylcytidine, 5-hydroxymethylcytidine, 5-carboxycytidine, 5-formylcytidine, 5-methoxycytidine, pseudouridine, 5-hydroxyuridine, 5-methyluridine, 5-hydroxymethyluridine, 5-carboxyuridine, 5-formyluridine, 5-methoxyuridine, 5-hydroxypseudouridine, 5-methylpseudouridine, 5-hydroxymethylpseudouridine, 5-carboxypseudouridine, 5-formylpseudouridine, and 5-methoxypseudouridine, optionally at an amount of at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or 100% of the non-canonical nucleotides. 
     
     
         73 . The method of  claim 65 , wherein the synthetic RNA comprises a 5′ cap structure. 
     
     
         74 . The method of  claim 65 , wherein the synthetic RNA comprises a 5′-UTR comprising a Kozak consensus sequence, the 5′-UTR optionally comprises a sequence that increases RNA stability in vivo, and the 5′-UTR optionally comprises an alpha-globin or beta-globin 5′-UTR. 
     
     
         75 . The method of  claim 65 , wherein the synthetic RNA comprises a 3′-UTR comprising a sequence that increases RNA stability in vivo, and the 3′-UTR optionally comprises an alpha-globin or beta-globin 3′-UTR. 
     
     
         76 . The method of  claim 65 , wherein the synthetic RNA comprises a 3′ poly(A) tail, optionally from about 20 nucleotides to about 250 nucleotides in length. 
     
     
         77 . The method of  claim 65 , wherein the prodrug is administered systemically. 
     
     
         78 . The method of  claim 65 , wherein the cancer is selected from the basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; lymphoma including Hodgkin's and non-Hodgkin's lymphoma, as well as B-cell lymphoma (including low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; as well as other carcinomas and sarcomas; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), and Meigs' syndrome. 
     
     
         79 .- 84 . (canceled) 
     
     
         85 . The method of  claim 65 , wherein the cell is a skin cell. 
     
     
         86 . The method of  claim 85 , wherein the skin cell is selected from the group consisting of a fibroblast, a keratinocyte, a melanocyte, an adipocyte, a mesenchymal stem cell, an adipose stem cell, and a blood cell. 
     
     
         87 . The method of  claim 65 , further comprising transfecting the cell with a synthetic RNA encoding a targeting molecule capable of directing the cell to a tumor, to result in the cell expressing the targeting molecule. 
     
     
         88 . The method of  claim 77 , wherein the prodrug is administered by injection.