中文摘要：本发明公开了一种能靶向RNA的Cas9多肽及其制备方法和应用。具体为一种基因工程核蛋白组合物，包括一个与Cas9多肽相关的规律成簇的间隔短回文重复（CRISPR），一个重组RNA或合成的单一导向RNA（sgRNA）。
 
外文摘要：Cas9 polypeptides which target RNA and methods of using them are provided.
NOVELTY - Engineered nucleoprotein complex comprises: a clustered regularly interspaced short palindromic repeats (CRISPR) associated protein 9 (Cas9) polypeptide; and a recombinant or synthetic single guide RNA (sgRNA).
USE - The engineered nucleoprotein complex is useful for treating or ameliorating a disease, condition, or infection in a mammalian or a human subject caused by a repeat sequence of ctg, cctg, cag and/or ggggcc, where the disease or condition is caused by or is associated with a RNA microsatellite repeat expansion, or is myotonic dystrophy, Huntington's disease, familial amyotrophic lateral sclerosis, cancer, spinal muscular atrophy, spinocerebellar ataxia, fragile X-associated tremor/ataxia syndrome, spinal-bulbar muscular dystrophy, oculopharyngeal muscular dystrophy or fragile X syndrome, and the infection is a viral or bacterial infection, preferably herpes viridae or herpes s! implex virus, a HIV, Epstein Barr virus, hepatitis virus, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, Zika virus, enteroviruses, human papilloma virus (HPV), influenza virus, mar-burg virus, Ebola virus, Mumps virus, Cytomegalovirus, rotavirus, Rubella virus, Varicella zoster virus, severe acute respiratory syndrome coronavirus, a Paramyxoviridae or measles virus, West Nile virus, Yellow fever virus, or Dengue fever virus infection (all claimed). Test details are described but no results given.
 
主权项：DETAILED DESCRIPTION - Engineered nucleoprotein complex comprises: a clustered regularly interspaced short palindromic repeats (CRISPR) associated protein 9 (Cas9) polypeptide; and a recombinant or synthetic single guide RNA (sgRNA). The Cas9 polypeptide: lacks all or part of an H-N-H domain, all or part of at least one RuvC nuclease domain, all or part of a Cas9 polypeptide DNase active site, and/or all or part of a beta beta alpha -metal fold comprisin! g a Cas9 polypeptide active site as compared to a correspondin! g wild t ype (WT) Cas9 polypeptide; lacks DNase and/or DNA cleaving capability or activity, or nickase activity, where the DNase or DNA cleaving capability or activity is removed by mutation or removal of all or part of an H-N-H domain, all or part of at least one RuvC nuclease domain of Cas9, and/or all or part of a Cas9 polypeptide DNase active site or a beta beta alpha -metal fold comprising a Cas9 polypeptide active site; and has a reduced polypeptide size that permits packaging of the Cas9-coding nucleotide in a viral or other delivery vector. The sgRNA is engineered or designed to comprise: on its 5' end, an RNA sequence that hybridizes to or binds to a target RNA; and (2) on its 3' end (i) an RNA sequence capable of binding to or associating with the Cas9 polypeptide and/or a Cas9 polypeptide binding scaffold sequence, or (ii) a linker that binds or covalently or non-covalently links the 5' RNA-hybridizing or binding end of the sgRNA with the Cas9 polypeptide. The sgRNA 3' end! or scaffold sequence comprises all or part of, or is derived from, the WT cognate guide nucleic acid of any one of the archaeal or bacterial Cas9 polypeptide. The nucleoprotein complex does not comprise a protospacer adjacent motif-presenting oligonucleotide and does not cleave or nick DNA or genomic loci. INDEPENDENT CLAIMS are also included for:
(1) treating or ameliorating a disease, condition, or infection in a mammalian or a human subject, comprising administering (a) the engineered nucleoprotein complex, or (b) a nucleic acid, nucleic acids, vector or vectors, encoding the engineered nucleoprotein complex, where the Cas9 polypeptide is adapted to be associated with, or that binds to or is covalently or non-covalently linked to, an effector polypeptide, and the nucleic acid is, or nucleic acids are, expressed intracellularly and express the engineered nucleoprotein complex to the mammalian or human subject; and
(2) a composition comprising the engineered nucleoprotein complex, or the nucleic acid or nucleic acids, vect or or vectors, encoding the engineered nucleoprotein complex, and an excipient.
 
申请号：US201615359567
公开/公告号：US2017145394A1
申请日：2016-11-22
公开/公告日：2017-05-25
申请/专利权人：UNIV CALIFORNIA (REGC-C); BATRA R (BATR-I); FANG M (FANG-I)
发明/设计人：BATRA, R; FANG, M; NELLES, D A; YEO, G; NELLES, D
分类号：A61K38/465; A61K48/0058; C07K19/00; C12N15/111; C12N15/113; C12N15/62; C12N9/22; C07K2319/01; C07K2319/85; C12N2310/10
主分类号：A61K38/46; A61K48/00; C12N9/22
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