公开(公告)号：US20120282225A1

公开(公告)日：2012-11-08

申请号：US13500010

申请日：2010-09-13

申请人： Samir El-Andaloussi ,  Gregory Heller ,  Ulo Langel ,  Taavi Lehto ,  Andres Merits ,  Liane Ulper

发明人： Samir El-Andaloussi ,  Gregory Heller ,  Ulo Langel ,  Taavi Lehto ,  Andres Merits ,  Liane Ulper

IPC分类号： A61K35/76 ,  A61P35/00 ,  C12N15/113

CPC分类号： A61K35/761 ,  A61K35/768 ,  C12N7/00 ,  C12N15/11 ,  C12N15/113 ,  C12N15/86 ,  C12N2310/11 ,  C12N2320/33 ,  C12N2710/10332 ,  C12N2710/10343 ,  C12N2710/10361 ,  C12N2770/36132 ,  C12N2770/36143 ,  C12N2770/36161 ,  C12N2840/44

摘要： Viral vectors are potential tools for eliminating the viability of eukaryotic cells in anti-cancer therapies since they can efficiently destroy the cancer cells and trigger an immune response against tumours. Typically viruses are not specific to cancer cells and all methods known in art aiming to the construction of cancer-specific viruses suffer from serious problems. The present invention presents a universal method to overcome these problems and is usable for any DNA virus replicating in nucleus or for any layered vector of RNA viruses. In this method the viral gene expression and/or replication will be blocked by the introduction of one or more aberrantly spliced introns into crucial gene expression units of the virus or vector. Lethal effect of these mutations is reverted in a controlled manner by the delivery of splice-switch oligonucleotide (s) correcting the introduced defects and restoring the biological functionality of the virus or vector, including cytolytic properties.

摘要翻译： 病毒载体是消除抗癌治疗中真核细胞活力的潜在工具，因为它们可以有效地破坏癌细胞并引发针对肿瘤的免疫应答。 通常，病毒对癌细胞不是特异性的，并且用于构建癌症特异性病毒的艺术中已知的所有方法都遭受严重问题。 本发明提出了克服这些问题的通用方法，并且可用于在核中或任何分层RNA病毒载体中复制的任何DNA病毒。 在该方法中，通过将一个或多个异常剪接的内含子引入病毒或载体的关键基因表达单元中，病毒基因表达和/或复制将被阻断。 通过递送剪接切换寡核苷酸来校正引入的缺陷并恢复病毒或载体的生物学功能，包括细胞溶解性质，这些突变的致死作用以受控的方式被还原。

公开(公告)号：US20110130304A1

公开(公告)日：2011-06-02

申请号：US12675953

申请日：2008-08-29

申请人： Kai Rauasalu ,  Anna Iofik ,  Valeria Lulla ,  Liis Karo-Astover ,  Kristi Tamm ,  Liane Ulper ,  Inga Sarand ,  Andres Merits

发明人： Kai Rauasalu ,  Anna Iofik ,  Valeria Lulla ,  Liis Karo-Astover ,  Kristi Tamm ,  Liane Ulper ,  Inga Sarand ,  Andres Merits

IPC分类号： C40B40/02 ,  C40B50/06 ,  C40B40/06 ,  C07H21/04 ,  C12N15/63

CPC分类号： C12N15/86 ,  C12N15/1027 ,  C12N15/1034 ,  C12N2770/36143

摘要： A method for creating an alphavirus-based genomic library, comprising a) ligation of foreign sequence (s) from an expression library or a random library into plasmids containing cloned alphaviral cDNA, b) multiplication of the obtained plasmid constructs in bacterial cells, c) direct transfection of the obtained plasmid constructs into mammalian or arthropod cells, characterized in that the sequence of an intron or sequences of introns are inserted into the respective genome of an alphavirus or into the cDNA of an expression vector based on an alphavirus, —the sequence of a viral subgenomic promoter, which is larger than minimal functional promoter is inserted immediately to the 3′ end of the sequences coding the structural proteins of the named alphavirus, —and ribozyme sequence is inserted for creating correct 3′ ends of the alphavirus.

摘要翻译： 一种用于产生基于甲病毒病毒的基因组文库的方法，包括：a）将外源序列从表达文库或随机文库连接到含有克隆的甲病毒cDNA的质粒中，b）将获得的质粒构建体在细菌细胞中增殖，c） 将获得的质粒构建体直接转染到哺乳动物或节肢动物细胞中，其特征在于将内含子或内含子序列的序列插入到甲病毒病毒的相应基因组或基于甲病毒属的表达载体的cDNA中，该序列 将大于最小功能性启动子的病毒亚基因组启动子立即插入到编码所述甲病毒属的结构蛋白的序列的3'末端，并插入核酶序列以产生甲病毒的正确3'末端。