Top > Search of International Patents > BORNA VIRAL VECTOR AND UTILIZATION THEREOF


Foreign code F180009589
File No. 5625
Posted date Nov 12, 2018
Country WIPO
International application number 2018JP008652
International publication number WO 2018168586
Date of international filing Mar 6, 2018
Date of international publication Sep 20, 2018
Priority data
  • P2017-049247 (Mar 14, 2017) JP
Abstract A viral vector comprising (a) a cDNA of a recombinant viral RNA having a sequence in which a foreign gene is inserted into a sequence carrying at least N gene, X gene, P gene and L gene of a Borna disease virus genome, said genes being arranged in the same order as in the Borna disease virus genome, (b) a DNA encoding a ribozyme, and (c) a promoter sequence, wherein (b) is located upstream and downstream of (a) and (a) and (b) are located downstream of (c), characterized in that (a) is a cDNA of a recombinant viral RNA in which G gene in the Borna disease virus genome is disrupted and the sequence of G gene of an avian Borna virus genome is also present. The present invention can be appropriately used, as a gene transfer technique not affecting the host chromosome, in various fields of, for example, treating and preventing brain nervous system diseases, techniques for visualizing neural cells in the brain neurology area, etc.
Outline of related art and contending technology BACKGROUND ART
Foreign genes in a biological or cell as a means of transport, has been known a method of using a viral vector.To this, a retrovirus, lentivirus, adenovirus, adeno-associated virus, using Sendai virus vectors have been developed.
However, these existing viral vector has, for example in the case of viral DNA into the host chromosome may be incorporated into the viral genes to the host (e.g., human) pathogenic expression was a problem.In addition, can be infected by virus host range is narrow, can be applied only to a particular organism or a problem, an exogenous gene into the viral genome integration site and by a difference between the gene introduction efficiency, the problem of gene introduction efficiency was poor.In addition, in vivo immune response or eliminated by the virus, viral mutations in the gene or promoter or can be due to a change of the efficiency, stability and also a problem of low persistence.
Further, such as in gene therapy, gene only cells of interest can be stored in the expected gene introduction technology.In particular, the treatment of diseases of the nervous system is considered to be effective for gene therapy, gene can be introduced into nerve cells selectively, further safety, stability, and high persistence of gene introduction efficiency has been desired the development of a viral vector.
Borna virus is, non-negative single-stranded segments of the chain has as its genome RNA (Mononegavirales) belonging to the virus and Mononegavirales, infected cells having characteristics of directivity.Current, borna virus family is borna virus (Bornaviridae) genus (Bornavirus), virus infected mammal borna (Borna disease virus: BDV) of the other, birds borna virus infected birds (Avian Bornavirus: ABV) have been identified and the like.
BDV replicating virus in the cell nucleus, the infection is a non-toxic and is characterized in that a long time and persistently, such as infection has a feature that extremely broad host range (such as 2 and non-patent document 1).
BDV is used as a technique for introduction of foreign genes into cells, and a green fluorescent protein (GFP) expression cassette 5 BDV genome ' untranslated region of the insertion end side, and the recombinant virus infected rats with high activity of the polymerase to, the GFP gene is expressed in the rat neurons has been reported (Non-Patent Document 3).
Is Patent Document 1, the viral genome (a) g cDNA encoding borna's translation of the foreign gene is inserted into an arbitrary area of a recombinant virus cDNA, cDNA (b) encode (c) and comprises a promoter sequence, the upstream and downstream of the (a) arranged (b), and (b) (a) and (c) is disposed downstream of the viral vectors is disclosed.
In addition, in Patent Document 2, the viral genome (a) at least in the N gene borna disease, gene X, P L gene and the gene, in the same order in which the viral genome borna has a disease, wherein the translation of the gene P untranslated region connected to the downstream region of the foreign gene is inserted into a recombinant virus RNA having a sequence of the cDNA, DNA (b) encode (c) a promoter sequence and, the upstream and downstream of the (a) arranged (b), and (b) (a) and (c) is disposed downstream of and is characterized in that the vector has been disclosed.
Scope of claims (In Japanese)[請求項1]








 請求項1~7のいずれかに記載のウイルスベクターと共に、ヘルパープラスミドとして、ボルナ病ウイルスゲノムのN遺伝子、P遺伝子、L遺伝子及び鳥ボルナウイルスゲノムのG遺伝子を発現するプラスミド又はプラスミド群をin vitroの細胞に導入する工程、ならびに該ウイルスベクター及びヘルパープラスミドを導入した細胞を培養して組換えウイルスを産生させる工程を含むことを特徴とする組換えウイルスの作製方法。

 請求項1~7のいずれかに記載のウイルスベクターと共に、ヘルパープラスミドとして、ボルナ病ウイルスゲノムのN遺伝子、P遺伝子及びL遺伝子を発現するプラスミド又はプラスミド群と鳥ボルナウイルスゲノムのG遺伝子を発現するプラスミドとをin vitroの細胞に導入する工程、ならびに該ウイルスベクター及びヘルパープラスミドを導入した細胞を培養して組換えウイルスを産生させる工程を含むことを特徴とする組換えウイルスの作製方法。

 ヘルパープラスミドとして、さらに、ボルナ病ウイルスゲノムのM遺伝子を発現するプラスミドをin vitroの細胞に導入する請求項9又は10に記載の組換えウイルスの作製方法。




  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • Inventor
  • TOMONAGA, Keizo
  • MAKINO, Akiko
IPC(International Patent Classification)
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