CN102676529A - 一种反义寡核苷酸 - Google Patents
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Abstract
本发明提供了一种反义寡核苷酸。在本发明中提供了使用外显子-居间AON来优化外显子跳跃的手段和方法。我们证实了通过靶向外显子中的推断的剪接调控序列(ESE)可提高跳跃效率。这种双重靶向对于在本发明之前难以获得有效跳跃的外显子是特别有益的。本发明还提供了表达所述反义寡核苷酸的载体。
Description
本申请是申请日为2007年5月18日的题为″用于诱导外显子跳跃的手段和方法″的中国专利申请No.200780018157.5的分案申请。
技术领域
本发明涉及寡核苷酸及其等同物以及将它们用于诱导外显子从mRNA中去除的应用。
背景技术
近年来,反义技术已经得到相当大的发展。反义技术的一般领域可分为若干方面。这些方面之一是外显子跳跃领域。
在该领域中,向细胞中加入小分子从而特异地干扰信使RNA(mRNA)的剪接,并且更具体地,干扰成熟的mRNA中的特定外显子的结合,或者更准确地说是成熟的mRNA中的特定外显子结合不足。
在外显子跳跃领域中,可以区分为两种一般方法。一种方法集中在干扰剪接(即,外显子序列的物理连接)的酶促过程。另一种方法依赖于干扰剪接过程的″逻辑学″(logistics),即干扰通过剪接机制对外显子的识别。两种方法均使用小分子来获得所需的效果。该小分子都共有核酸分子能够″杂交″至互补序列的特性。这样的小分子的实例当然是RNA和DNA、以及修饰的RNA和DNA(如LNA和吗啉代化合物(morpholinos)、以及诸如PNA的核酸模拟物)。这些分子进一步被称为通用术语″小分子″或更加经常使用的术语″反义寡核苷酸(AON)″。
小分子被认为是通过杂交至pre-mRNA(前体mRNA)的特定位点而发挥作用的。杂交被认为干扰剪接的酶促过程或者外显子的识别。
目前,外显子跳跃集中在将一种或多种外显子从成熟的mRNA中特异性去除。然而,使剪接机制改变方向从而将其他的外显子包括在mRNA中也是可能的。例如,这种方法可应用在进行选择性剪接的基因中。在这些情形下,可将小分子设计为使剪接机制改变方向,以便更加有效地形成具体的可选择方案(specific alternative)。
发明内容
本发明提供一种反义寡核苷酸(AON),包括表2中示出的序列或者由表2中示出的序列组成。
根据本发明的AON,在一种实施方式中,包括14-40个核苷酸或其类似物。
根据本发明的AON,在一种实施方式中,包括14-25个核苷酸或其类似物。
根据本发明的AON,在一种实施方式中,所述AON包括SEQID NO:118表示的h44AON4或者由SEQ ID NO:118表示的h44AON4组成。
根据本发明的AON,在一种实施方式中,所述AON是修饰的寡核苷酸。
根据本发明的AON,在一种实施方式中,所述AON包括至少一个核苷酸类似物。
根据本发明的AON,在一种实施方式中,所述AON包括选自2′-O-甲基核糖核苷酸、硫代膦酸酯骨架、吗啉代核苷酸、锁核酸(LNA)核苷酸、和/或乙撑桥联核酸(ENA)核苷酸中的一种或多种修饰。
根据本发明的AON,在一种实施方式中,所述AON包括一个或多个2′-O-甲基寡核糖核苷酸、硫代膦酸酯骨架、吗啉代寡核苷酸、锁核酸(LNA)寡核苷酸、和/或乙撑桥联核酸(ENA)寡核苷酸。
根据本发明的AON,在一种实施方式中,所述AON包括吗啉代磷酰二胺寡核苷酸。
根据本发明的AON,在一种实施方式中,所述AON包括一个或多个2′-O-甲基硫代膦酸酯核苷酸。
根据本发明的AON,在一种实施方式中,所述AON包括一个或多个2′-O-甲基硫代膦酸酯寡核苷酸。
根据本发明的AON,在一种实施方式中,包括全长硫代膦酸酯骨架,且其中,所有核苷酸包括2′-O-甲基修饰。
根据本发明的AON,在一种实施方式中,少于10%的核苷酸是核苷酸类似物。
本发明还提供了一种载体,表达上述AON或其前体。
根据本发明的载体,在一种实施方式中,其为病毒载体。
根据本发明的载体,在一种实施方式中,其为腺相关病毒载体。
在本发明中,已经发现外显子跳跃过程能够变得更加有效。许多关于外显子跳跃的早期研究是使用针对抗肌萎缩蛋白基因中外显子跳跃的AON而进行的。迄今为止,我们已经识别了一系列的114个外显子-居间AON(exon-internal AON),它们在培养的肌细胞中诱导35个外显子的跳跃(14)。我们的数据表明,有效的外显子-居间AON可能是通过对富含丝氨酸/精氨酸的(SR)蛋白家族成员结合至外显子剪接增强子(ESE)位点的空间位阻而发挥作用的(15)。SR蛋白结合至外显子序列移动(exon sequence motive)并集结其他的剪接因子以介导剪接作用(19)。对于我们所研究的多数外显子而言,单个ESE的AON-靶向(AON-targeting)足以诱导显著水平的外显子跳跃(14)。然而,对于一些外显子而言,跳跃效率低至0。在本发明中,我们证实了通过用两个或多个外显子居间AON来靶向外显子可以提高跳跃水平。为此目的,本发明提供了一种用于确定在产生含所述mRNA前体的外显子的细胞中是否能提高从成熟的mRNA中去除外显子的效率的方法,
所述方法包括:
为所述细胞提供能够杂交至所述外显子的外显子居间部分的第一反义寡核苷酸(AON),并在为所述细胞提供能够杂交至所述外显子的另一外显子居间部分的第二AON后,确定所述效率是否得到提高。优选通过与每一种单独AON的效率比较水平来确定效率的提高。本文使用的术语″提高的效率″是指这样的情形,其中(i)去除了靶标外显子的mRNA的量高于每一种单独AON,或(ii)在所述细胞中能够检测靶标外显子的去除的时间间隔增加,或(iii)二者的结合。与使用单个AON的外显子跳跃方法相比,本发明的方法还提供了额外的稳健性。因此,本发明进一步提供了一种用于在由细胞产生的pre-mRNA中诱导外显子跳跃的方法,该方法包括为所述细胞提供至少两个能够杂交至所述pre-mRNA上外显子的不同外显子居间部分的AON,并培养所述细胞以允许所述pre-mRNA进行剪接。
一种序列由于其终止于成熟mRNA中的pre-mRNA的一部分,因而通常被称为外显子。pre-mRNA包含至少一个内含子。第一个外显子包含帽子-位点而最后一个外显子通常包含poly A信号。在本发明中,外显子跳跃优先定向于居间外显子。居间外显子是外显子2至外显子n-1中的任一个,其中n是所述pre-mRNA中的外显子总数。当外显子跳跃时,其就不再包括在成熟mRNA中。这种特征可被一些人看作是,预示着该序列不再是外显子而是内含子(或内含子的一部分)。因此,在本文中将外显子限定为存在于pre-mRNA中的序列,在不为细胞提供诱导所述外显子跳跃的AON时,其通过表达所述pre-mRNA的细胞的剪接机制而结合至成熟的mRNA中。因此,本发明中的外显子跳跃方法也被认为是用于将外显子序列改变为内含子序列的方法,可替换地,本发明的方法是一种用于掩蔽(masking)外显子序列的方法,因而当外显子序列被掩蔽时,所述外显子序列不再被细胞的剪接机制识别为外显子。
可以以多种不同的方式来为细胞提供AON。在一种优选的实施方式中,通过将所述AON结合至基因递呈载体(优选脂质体)中从而将所述AON提供至所述细胞中。在另一种优选的实施方式中,通过表达所述AON或其前体的载体的方式将所述AON提供至所述细胞中。在一种优选的实施方式中,所述载体包括病毒载体、优选腺相关病毒载体。因此,本发明进一步提供一种基因递呈载体,包括至少两个对相同外显子特异的外显子居间AON。
使用至少两个对相同外显子特异的外显子居间AON使得本发明的方法更加强有力。此外,其使得当仅使用一个AON时不能有效跳跃的外显子有效跳跃。不受理论的限制,可以认为外显子-居间AON通过阻遏结合至外显子剪接增强子(ESE)位点的所谓的SR(Ser-Arg)蛋白的结合来诱导外显子的跳跃。在本发明中,除此之外,发现外显子可包含所谓的独立ESE位点。这意味着当一个ESE位点的活性由于例如所述外显子上的AON的存在而被阻断或阻遏时,在所述外显子上的另一个独立的ESE仍保持功能性。由于该ESE保持功能性,该外显子仍然有效地结合至mRNA中。这种效应可以通过靶向所述外显子上的所有独立的ESE位点而被抵消。在这种情况下,基本上所有的ESE位点被AON靶向,从而阻断或阻遏SR蛋白结合至所述ESE位点,所述ESE位点随后导致由此靶向的外显子的有效跳跃。可以以多种方式来实现对ESE位点的阻断或阻遏。例如,所述AON的杂交可改变外显子的二级结构,因而使SR蛋白不再结合至所述ESE位点。在一种优选的实施方式中,所述AON与靶标外显子上预测ESE位点的至少部分重叠。这样,SR蛋白与所述位点的结合由于所述AON与其杂交而受到位阻。优选所述AON与至少一个预测ESE位点重叠。当预测的ESE位点具有成簇的倾向时,所述AON优选与所述外显子上的至少两个、优选至少三个预测ESE位点重叠。因此,在一种优选的实施方式中,所述第一AON在所述外显子RNA上的杂交位点与所述外显子RNA上的至少一个预测ESE位点重叠。优选地,所述第二AON在所述外显子RNA上的杂交位点与所述外显子RNA上的至少一个预测ESE位点重叠。
将所述第一AON和所述第二AON靶向于所述外显子RNA上的单个预测ESE位点并不是优选的。尽管可能有这样的情况,优选的是所述第一AON靶向于不同的ESE而不是所述第二AON。这可以以多种方式实现。优选通过避免所述AON在所述外显子RNA上杂交位点的显著重叠来避免对相同ESE的靶向。因此,优选所述第一AON的杂交位点与所述第二AON的杂交位点表现为少于5个连续核苷酸重叠。更加优选地,所述位点具有少于3个并且更加优选少于1个重叠或没有重叠。可以容易地通过AON的选择来避免这样的重叠。由于本发明的AON优选在所述外显子RNA上具有线性且连续的杂交位点,因而优选通过避免所使用的AON中具有显著的序列同一性而避免重叠。在这个方面,优选所述第一AON和所述第二AON具有少于5个连续的核苷酸序列同一性。优选在所述外显子RNA上的所述第一和所述第二杂交位点具有少于3个连续的核苷酸重叠、更加优选少于2个、并且优选少于1个核苷酸重叠或没有重叠。类似地,所述第一AON和所述第二AON优选具有少于3个连续的核苷酸序列同一性、更加优选少于2个并且优选少于1个核苷酸序列同一性。在一种优选的实施方式中,所述外显子包括至少2个独立的ESE位点。所述第一AON优选靶向于至少2个独立的ESE位点而所述第二AON靶向于其中的第二个。
本文中使用的限定条件″序列同一性(sequence identity)″优选指应用核苷酸A、C、G或T的同一性,其中T可由U替代。近年来,已经产生许多不同的核苷酸类似物。其中的一些与所描述的典型或天然核苷酸在种类和数量上具有相同的结合特性。然而,它们中的大多数在种类上具有相似的(mimic)结合特性,而在数量上却没有。在许多情况下,类似物与互补核苷酸的结合强度低于相应的经典核苷酸(classical nucleotide)。还有一些类似物与2个或多个经典核苷酸具有相似的结合特性,并且还有一些核苷酸类似物,它们被有效地结合于核酸中却不表现出对相对链上特定的经典核苷酸的显著选择性或不表现为结合至相对链上特定的经典核苷酸上。本领域中的技术人员熟知这些以及相似的类似物的细节并且能够将它们应用在本发明的AON中,只要该AON能够杂交至外显子RNA上的杂交位点即可。
本发明的AON优选包括14-40个核苷酸或其类似物。优选地,所述AON包括14-25个核苷酸或其类似物。本发明的AON优选包括少于10%的核苷酸类似物。AON优选包括少于4个核苷酸类似物、优选少于3个、更加优选少于2个。
本发明的AON优选为修饰的寡核苷酸。在一种尤其优选的实施方式中,所述AON包括一个或多个2′-O-甲基寡核糖核苷酸,这使得AON能够抵抗核糖核酸酶H诱导的DNA/RNA杂交链(hybrid)的降解。此外,可以利用硫代膦酸酯骨架(phosphorothiatebackbone)来提高AON对核酸酶的稳定性并增强细胞的摄取(cellular uptake)。在一种优选的实施方式中,本发明的AON具有全长硫代膦酸酯骨架并且所有碱基(核苷酸)具有2′-O-甲基修饰。可替换地,已经利用吗啉代磷酰二胺DNA(吗啉代化合物)、锁核酸(LNA)以及乙撑桥联核酸(ENA,ethylene bridged nucleic acids)寡聚物来调节剪接。因此,本发明的AON还可具有这些修饰。这些修饰使得寡聚物对核糖核酸酶H和核酸酶具有抗性并且提高其对于靶标RNA的亲和力。对于ENA和LNA修饰,这种提高是通过降低的序列特异性而实现的。
外显子的外显子居间部分,是不表现为与外显子的5′-或3′-末端重叠的部分。外显子的5′-或3′-末端包含与外显子连接区相关的序列信息。外显子的外显子居间部分是居间于外显子边界内的至少1个、以及至少2个、并且更加优选至少5个核苷酸。除了如上文所限定的外显子居间部分之外,外显子居间AON还可以具有延伸至外显子边界或超过边界的核苷酸。然而,这并不是优选的情况。
本发明的方法优选进一步包括,当为所述细胞提供所述第二AON可提高从产生所述mRNA前体的细胞中的所述成熟mRNA中去除所述外显子的效率时,将所述第一AON和所述第二AON包含在一组AON中。随后优选将所述组用于获得表达所述pre-mRNA的靶细胞中的外显子跳跃。本发明还提供了包括至少2个AON的组,其中所述AON的至少第一个能够杂交至外显子的外显子居间部分而所述AON的第二个能够杂交至所述外显子的另一个外显子居间部分。在本发明的一种优选实施方式中,所述组是通过本发明的方法获得的。在一种尤其优选的实施方式中,所述AON的所述第一个和所述第二个靶向于所述外显子RNA上的ESE位点。更加优选地,所述AON的所述第一个和所述第二个靶向于所述外显子RNA上的独立的ESE位点。
本发明的组可以以多种方式进行使用。在一种实施方式中,所述组用来至少部分地减少细胞中异常蛋白(aberrant protein)的产生。这样的蛋白可以是,例如肿瘤蛋白或病毒蛋白。在许多肿瘤中,不仅存在肿瘤蛋白而且其相对的表达水平与肿瘤细胞的表型有关。类似地,不仅存在病毒蛋白而且在细胞中病毒蛋白的量均决定着特定病毒的毒性。此外,对于病毒的有效增殖和蔓延,在生命周期中的表达周期(timing of expression)以及细胞中某些病毒蛋白的量的平衡决定着病毒能否有效地产生。使用本发明的组,可以降低细胞中异常蛋白的量,使得例如肿瘤细胞的致瘤性(转移)减少和/或使得病毒侵染的细胞产生较少的病毒。
在一种优选的实施方式中,本发明的组用来将所述异常蛋白转化为功能蛋白。在一种实施方式中,所述功能蛋白能够发挥正常存在于细胞中但不存在于待处理细胞中的蛋白的功能。即使是功能部分恢复也常常会导致这样处理的细胞性能显著改善。由于具有更好的性能,这样的细胞还可比未修饰的细胞具有选择性优势,因而有助于处理的有效性。
本发明的这个方面特别适用于缺陷基因表达的恢复。这是通过导致靶标外显子的特异性跳跃,从而绕开或校正有害突变(通常为导致翻译终止的终止突变或移码点突变、单-或多-外显子缺失或插入)而实现的。
与基因-导入策略(gene-introduction strategy)相比,外显子跳跃基因理论要求给予少得多的治疗剂,通常为,但不限于,包括或类似于约14-40个核苷酸的AON。在一种优选的实施方式中,使用14-25个核苷酸的AON,因为这些分子更易于产生并可更加有效地进入细胞。本发明的组使对于随后的有效且安全的外显子跳跃策略和/或给药系统的设计具有更多的灵活性。基因功能恢复AON组的另外一个重要的优势是其恢复(至少是一些)内源基因活性,其仍进行它的大多数或所有基因调控线路(gene-regulatory circuitry),从而确保适当的表达水平和组织特异性异构体的合成。
本发明的这个方面原则上可应用于任何遗传病或疾病的遗传易感性,其中,当翻译读码框被原始突变打断时,特定外显子的靶向性跳跃可恢复翻译读码框。当内部稍短或改变的蛋白的翻译具有全部或部分功能的时候,这种应用尤其有益。优选的实施方式(对于该优选实施方式,这种应用可能具有治疗价值)是:对于肿瘤抑制基因(例如在乳腺癌、结肠癌、结节性硬化症、神经纤维瘤等中涉及的那些)中第二命中突变的素因(predisposition),其中活性的(部分)恢复通过第二命中突变将消除缺对染色体(缺体型,nullosomy)的现象,并因此将防止肿瘤发生。另一种优选的实施方式涉及具有直接导致疾病发生作用的缺陷型基因产物的(部分)恢复,例如,血友病A(凝血因子VIII缺乏)、先天性甲状腺功能低下症的一些形式(由于甲状腺球蛋白合成缺乏)以及杜兴氏肌营养不良(Duchenne Muscular Dystrophy,DMD)(其中伴X染色体的肌营养不良蛋白基因中的移码缺失、复制以及终止突变导致严重的、进行性肌退化)。DMD在青春期晚期或成人期早期通常是致命的,而相同基因中的非移码缺失或复制则会导致轻微得多的贝克尔氏肌营养不良(Becker Muscular Dystrophy,BMD),其与35-40岁至正常的预期寿命相一致。在正如应用于DMD的实施方式中,本发明通过如所需要的多个邻近的外显子而使外显子跳跃能够延及存在的缺失(或改变存在复制的mRNA产物),从而恢复读码框并产生内部稍微变短但仍具有功能的蛋白。基于同样(equivalent)具有这种诱导缺失的BMD患者具有轻微得多的临床症状,DMD患者的病情在AON-治疗后将有一个轻微得多的过程。考虑到本发明的组的许多临床应用,本发明的方法优选进一步包括向人类细胞提供所述组。在一种实施方式中,本发明提供了可通过本发明的方法获得的一组AON。在一种优选的实施方式中,所述组包括表1或表2中的AON中的至少一种。在一种优选的实施方式中,本发明提供了包括至少表1和/或表2中的AON的一组AON。在这种实施方式的一个优选方面,表1和/或表2中的至少2个AON的所述组包括h2AON1和h2AON3、h43AON1和h43AON2、h43AON1和h43AON3、h43AON1和h43AON4、h43AON1和h43AON5、h43AON2和h43AON5、h43AON3和h43AON5、h43AON4和h43AON5、h45AON1和h45AON4、h45AON1和h45AON5、h45AON1和h45AON9、h45AON2和h45AON3、h45AON2和h45AON4、h45AON2和h45AON5、h45AON3和h45AON4、h45AON3和h45AON5、h45AON3和h45AON9、h45AON4和h45AON5、h45AON5和h45AON9、h46AON4和h46AON8、h46AON4和h46AON20、h46AON4和h46AON22、h46AON6和h46AON21、h46AON8和h46AON22、h46AON8和h46AON23、h46AON8和h46AON26、h46AON9和h46AON21、h46AON9和h46AON22、h46AON9和h46AON26、h46AON22和h46AON24、h46AON24和h46AON26、h47AON1和h47AON3、h47AON1和h47AON5、h47AON2和h47AON3、h47AON2和h47AON5、h47AON3和h47AON4、h47AON3和h47AON5、h47AON3和h47AON6、h47AON4和h47AON5、h47AON5和h47AON6、h48AON 1和h48AON4、h48AON1和h48AON6、h48AON1和h48AON8、h48AON1和h48AON9、h48AON1和h48AON10、h48AON2和h48AON10、h48AON3和h48AON4、h48AON4和h48AON6、h48AON6和h48AON8、h48AON6和h48AON9、h48AON6和h48AON10、h48AON7和h48AON9、h57AON1和h57AON3和/或h57AON2和h57AON3。在一种优选的实施方式中,所述表1和/或表2中的至少2个AON的组包括h43AON1和h43AON3、h45AON1和h45AON4、h45AON1和h45AON5、h45AON1和h45AON9、h45AON2和h45AON3、h45AON2和h45AON4、h45AON3和h45AON4、h45AON3和h45AON5、h45AON3和h45AON9、h45AON4和h45AON5、h45AON5和h45AON9、h46AON9和h46AON21、h47AON1和h47AON3、h47AON1和h47AON5、h47AON2和h47AON3、h47AON2和h47AON5、h47AON3和h47AON4、h47AON3和h47AON5、h47AON3和h47AON6、h47AON4和h47AON5、h47AON5和h47AON6、h48AON1和h48AON4、h48AON1和h48AON8、h48AON3和h48AON4、h57AON1和h57AON3、h57AON2和h57AON3。在一种尤其优选的实施方式中,所述表1和/或表2中的至少2个AON的组包括h45AON1和h45AON4、h45AON1和h45AON5、h45AON1和h45AON9、h45AON2和h45AON3、h45AON3和h45AON4、h45AON3和h45AON5、h45AON3和h45AON9、h45AON4和h45AON5、h45AON5和h45AON9、h47AON1和h47AON3、h47AON1和h47AON5、h47AON2和h47AON3、h47AON2和h47AON5、h47AON3和h47AON4、h47AON3和h47AON5、h47AON3和h47AON6、h47AON4和h47AON5、h47AON5和h47AON6、h57AON1和h57AON3、h57AON2和h57AON3。本发明进一步提供了表2的AON。优选地,表2中的所述AON是h33AON1、h33AON2、h44AON3、h44AON4、h45AON11或h52AON3。
在一个方面,本发明进一步提供了用于治疗患有肿瘤、病毒感染和/或遗传疾病和/或遗传易感性的个体的方法,该方法包括对所述个体给予本发明的AON组。本发明进一步提供了一种包括根据本发明的AON组的药物组合物。优选地,所述组用于治疗患有肿瘤、病毒感染和/或遗传疾病和/或遗传易感性的个体。优选地,所述组用于治疗杜兴氏肌营养不良(DMD)。
本发明还提供了一种产生含有mRNA前体的外显子的细胞,其中,所述细胞包括能够杂交至所述外显子的外显子居间部分的第一AON、以及能够杂交至所述外显子的另一外显子居间部分的第二AON。
具体实施方式
实施例
结果与讨论
在一个外显子中的双重靶向
我们假设通过在一个外显子中靶向多个ESE,对于使用单独的AON不能以高水平跳跃的外显子,可以提高跳跃效率。图1A中描述了对于这些中的每个外显子的AON与推断的ESE(由ESEfinder预测得到的)的相对位置。
我们首先测试了对2个不可跳跃的外显子(外显子47和57)的双重靶向。当用这些AON的不同组合对人对照肌管培养物进行处理时,正如用RT-PCR分析所确定的(图1BC,附加表1),可获得外显子47和57的显著跳跃水平。有趣的是,对于外显子47,只有非重叠AON的组合诱导外显子跳跃,而那些重叠的组合则是无效的(附加表1)。类似地,对于外显子57,几乎重叠的AON的组合(h57AON1和h57AON2)不诱导外显子跳跃。这符合我们的假设,即这些外显子中存在两个相互排斥的ESE位点。
对于外显子2和外显子45,只有单个AON可重复性地诱导低水平的外显子跳跃(分别为,h2AON1和h45AON5)。可以通过将有效的h45AON5与无效的AON组合,以及通过结合2个单独的无效AON(例如h45AON3和h45AON9)来诱导外显子45的相当高水平的跳跃(图1D)。因此,对于这个外显子,也可能存在两个有效的ESE。相对而言,AON的组合不会提高外显子2的跳跃效率。实际上,将有效的h2AON1与无效的h2AON2组合消除了外显子2的跳跃(图1E)。对于h2AON1与h2AON3(重叠少于h2AON2)的组合却没有观察到这种干扰。对于外显子43和48,有效的AON只有到中等水平的跳跃。对于这些外显子,跳跃水平不会由于使用AON的组合而得到提高(附加表1)。在这些外显子中可以存在3个或甚至更多的ESE。因此,我们用3个不同AON的组合靶向于外显子48,仍然不能提高跳跃水平。因此,看起来这个外显子的剪接更加可能在很大程度上独立于ESE位点。这得到了以下事实的支持:预测的剪接位点是正确的(perfect)(3′剪接位点)或接近正确的(5′剪接位点),并且对于外显子48预测只有少量的ESE。
最后,我们使用外显子46特异性的AON组合,其中的一些单独使用就已经非常有效。我们目的是将跳跃水平提高至(接近)100%。然而,当与单个靶向相比时,所使用的组合中没有一种可进一步提高跳跃水平(附加表1)。这表明阻断1个ESE就足以扰乱该外显子的正确剪接。该外显子上的ESE位点彼此依赖,因而通过阻断一个,就可以使多个或所有ESE失活。可替换地,pre-mRNA的二级结构在结合AON后可发生改变,从而对于SR蛋白结合不再可获得其他的ESE位点。
有趣的是,不管单个AON是否有效,(部分)重叠组合看起来再次消极地干扰单独外显子的跳跃能力。预期2个无效的、重叠AON的组合也会是无效的,因为它们二者中的每一个靶向于非功能性ESE位点或其二者均靶向于相同的两个(或多个)相互排斥的ESE位点。对于重叠的、有效AON的组合,这种发现是未预料到的。然而,这些AON会彼此竞争,并迫使彼此从动态过程(dynamicprocess)中的靶标转录本中分离,从而对于SR蛋白可再次获得靶标位点。在结合后,SR蛋白将其他的剪接因子集结至剪接位点,并因而增强外显子的增加而非外显子跳跃。
材料与方法
AON和引物
上文中已经描述了所有用于双重靶向实验的AON(见附加表1)(11、14、15)。所有AON均包含2-O-甲基RNA、硫代磷酸酯骨架以及5′荧光素标记并且由Eurogentec(比利时)制造。(RT-)PCR引物选自跳跃的外显子侧面的外显子(Eurogentec,比利时;如果要求可提供序列)。
组织培养,转染以及RT-PCR分析
如上所述获得对照和DMD患者来源的肌管培养物(9、22)。对于外显子跳跃实验,以200nM的每种AON进行转染,至少进行两次。在所有的实验中,根据制造商的说明书,以每μg AON中2至3.5μl PEI使用聚乙烯亚胺(PEI,ExGen 500,MBI Fermentas)。取决于可用于每个靶标外显子的有效AON的数量,测试AON的不同组合(附加表1)。制备每种AON的AON-PEI配合物的分离溶液(separate solution)。基于核荧光信号的存在,转染效率通常超过80%。在转染后24-48小时分离RNA并如上所述进行RT-PCR分析(14)。使用靶标外显子侧面的外显子中的引物,通过RT-PCR分析来证实每种AON的成功转染。从琼脂糖凝胶中分离PCR片断并如上所述进行序列分析(10)。
表1.所使用的AON的特性
1++在正常的对照肌管培养物中,超过25%的转录本中观察到的外显子跳跃;+在达25%的转录本中观察到的外显子跳跃;-未检测到外显子跳跃。
2给出每一种AON对于每一种SR蛋白的最高值
3核苷酸中5′和3′剪接位点(SS)之间的核苷酸数量。3′和5′剪接位点之间的距离由靶标序列中第一个(3′剪接位点)或最后一个(5′剪接位点)核苷酸决定。
4在预测的二级RNA结构中AON靶向的可获得的核苷酸占AON全长的比例。
5在缺失Kobe中缺失的ESE中的AON靶标部分(Matsuo等,1990;Matsuo等,1991)
6先前已公开(van Deutekom等,2001)
7先前已公开(Aartsma-Rus等,2002)
8先前已公开(van Deutekom等,2001;Aartsma-Rus等,2003;Aartsma-Rus等,2004)
表2
附加表1.使用的AON的综述以及双重靶向实验所获得的结果
1″+″该单独的AON观察到外显子跳跃;″-″该单独的AON未(可重复地)观察到外显子跳跃
2与单靶向相比的双重靶向的结果;″+″双重靶向比用任何单个AON靶向更加有效;″=″双重靶向的有效性相当于最有效的单个AON;″-″双重靶向不必最有效的单个AON更有效或比其低效。
有效的AON为浅灰色阴影,比最有效的单个AON显著更好的组合为深灰色阴影,重叠组合加下划线。
附图说明
图1.双重靶向。
(A)在外显子2、43、45-48以及57中AON与推断ESE位点的相对位置。对于每个外显子,将外显子和AON按比例绘制为直线。通过ESEfinder预测得到的推断结合位点SF2/ASF、SC35、SRp40以及SRp55在其各自的位置上绘制为方框。
(B-E)双重靶向实验后RT-PCR分析的一些实例。
(B、C)对于外显子47和57,没有可应用的AON可重复地诱导外显子跳跃。然而,使用这些AON的组合,能够以显著水平可重复地诱导外显子47和57跳跃。对于外显子57,包含h57AON3的组合最为有效,而对于外显子47,所有非重叠的组合诱导相当水平的外显子47跳跃。在一些情况下,可在外显子47的PCR中观察到额外的条带,其比野生型产物略短。在经处理和未经处理的样品中均观察到该条带是不可复制的,并且似乎是含DMD外显子72-74的特异性PRC产物。
(D)对于外显子45,只有一个可应用的AON可重复地诱导跳跃,尽管是以低水平诱导(h45AON5)。对于h45AON1和h45AON4,偶尔可观察到极低水平的外显子跳跃,但却是不可重复的。使用AON的组合可以高得多的水平实现外显子45跳跃。对于h45AON5和h45AON1或h45AON3的组合以及h45AON1和h45AON9的组合,观察到最高水平的外显子45跳跃。相比之下,重叠的h45AON2和h45AON9的混合物是无效的。
(E)对于外显子2,只有重叠的AON可应用。当有效的h2AON1与无效的、重叠h2AON2组合时,不能诱导跳跃。当h2AON1与无效的、较少重叠的h2AON3组合时,没有观察到这种效应。Aartsma-Rus A等(2005)的论文中给出了所述AON的序列。114个外显子-居间AON对于靶向的DMD外显子跳跃的功能分析:对SR蛋白结合位点的空间位阻的指征。寡核苷酸15:284-297.NT是未转染的,-RT是阴性对照,M是100bp大小的标记(marker)。
参考文献
1.Emery AE.(2002).The muscular dystrophies.Lancet 359:687-95.
2.Monaco AP,Bertelson CJ,Liechti-Gallati S,Moser H,and KunkelLM.(1988).An explanation for the phenotypic differencesbetween patients bearing partial deletions of the DMD locus.Genomics 2:90-95.
3.Hoffman EP,et al.(1988).Characterization of dystrophin inmuscle-biopsy specimens from patients with Duchenne′s orBecker′s muscular dystrophy.N Engl J Med 318:1363-1368.
4.Koenig M,Hoffman EP,Bertelson CJ,Monaco AP,Feener C,andKunkel LM.(1987).Complete cloning of the Duchenne musculardystrophy(DMD)cDNA and preliminary genomic organization ofthe DMD gene in normal and affected individuals.Cell 50:509-517.
5.Yoshida M,and Ozawa E.(1990).Glycoprotein complexanchoring dystrophin to sarcolemma.J Biochem(Tokyo)108:748-752.
6.Hoffman EP,Brown RH,Jr.,and Kunkel LM.(1987).Dystrophin:the protein product of the Duchenne muscular dystrophy locus.Cell 51:919-928.
7.Lu QL,et al.(2003).Functional amounts of dystrophin producedby skipping the mutated exon in the mdx dystrophic mouse.NatMed 8:1009-1014.
8.van Deutekom JC,and van Ommen GJ.(2003).Advances inDuchenne muscular dystrophy gene therapy.Nat Rev Genet 4:774-783.
9.Aartsma-Rus A,et al.(2003).Therapeutic antisense-induced exonskipping in cultured muscle cells from six different DMD patients.Hum Mol Genet 12:907-914.
10.van Deutekom JC,et al.(2001).Antisense-induced exon skippingrestores dystrophin expression in DMD patient derived musclecells.Hum Mol Genet 10:1547-1554.
11.Aartsma-Rus A,et al.(2004).Antisense-induced multiexonskipping for duchenne muscular dystrophy makes more sense.AmJ Hum Genet 74:83-92.
12.Bremmer-Bout M,et al.(2004).Targeted Exon Skipping inTransgenic hDMD Mice:a Model for Direct Pre-clinicalScreening of Human-specific Antisense Oligonucleotides.Molecular Therapy 10:232-240.
13.Lu QL,et al.(2005).Systemic delivery of antisenseoligoribonucleotide restores dystrophin expression in body-wideskeletal muscles.Proc.Natl.Acad.Sci.U.S.A 102:198-203.
14.Aartsma-Rus A,Bremmer-Bout M,Janson A,Den Dunnen J,vanOmmen G,and van Deutekom J.(2002).Targeted exon skipping asa potential gene correction therapy for Duchenne musculardystrophy.Neuromuscul Disord 12:S71-S77.
15.Aartsma-Rus A,et al.(2005).Functional analysis of 114exon-internal AONs for targeted DMD exon skipping:indicationfor steric hindrance of SR protein binding sites.Oligonucleotides15:284-297.
16.Stojdl DF,and Bell JC.(1999).SR protein kinases:the splice oflife.Biochem Cell Biol 77:293-298.
17.Fokkema IF,den Dunnen JT,and Taschner PE.(2005).LOVD:easy creation of a locus-specific sequence variation database usingan″LSDB-in-a-box″approach.Hum.Mutat.26:63-68.
18.England SB,et al.(1990).Very mild muscular dystrophyassociated with the deletion of 46% of dystrophin.Nature 343:180-182.
19.Mirabella M,et al.(1998).Giant dystrophin deletion associatedwith congenital cataract and mild muscular dystrophy.Neurology51:592-595.
20.Bushby KM,Appleton R,Anderson LV,Welch JL,Kelly P,andGardner-Medwin D.(1995).Deletion status and intellectualimpairment in Duchenne muscular dystrophy.Dev Med ChildNeurol 37:260-269.
21.Tennyson CN,Klamut HJ,and Worton RG.(1995).The humandystrophin gene requires 16 hours to be transcribed and iscotranscriptionally spliced.Nat Genet 9:184-190.
22.Havenga MJ,et al.(2002).Exploiting the natural diversity inadenovirus tropism for therapy and prevention of disease.J Virol76:4612-4620.
Claims (16)
1.一种反义寡核苷酸(AON),包括表2中示出的序列或者由表2中示出的序列组成。
2.根据权利要求1所述的AON,包括14-40个核苷酸或其类似物。
3.根据权利要求2所述的AON,包括14-25个核苷酸或其类似物。
4.根据权利要求1至3中任一项所述的AON,其中,所述AON包括SEQ ID NO:118表示的h44AON4或者由SEQ ID NO:118表示的h44AON4组成。
5.根据权利要求1至3中任一项所述的AON,其中,所述AON是修饰的寡核苷酸。
6.根据权利要求1至3中任一项所述的AON,其中,所述AON包括至少一个核苷酸类似物。
7.根据权利要求5所述的AON,其中,所述AON包括选自2′-O-甲基核糖核苷酸、硫代膦酸酯骨架、吗啉代核苷酸、锁核酸(LNA)核苷酸、和/或乙撑桥联核酸(ENA)核苷酸中的一种或多种修饰。
8.根据权利要求5所述的AON,其中,所述AON包括一个或多个2′-O-甲基寡核糖核苷酸、硫代膦酸酯骨架、吗啉代寡核苷酸、锁核酸(LNA)寡核苷酸、和/或乙撑桥联核酸(ENA)寡核苷酸。
9.根据权利要求8所述的AON,其中,所述AON包括吗啉代磷酰二胺寡核苷酸。
10.根据权利要求5所述的AON,其中,所述AON包括一个或多个2′-O-甲基硫代膦酸酯核苷酸。
11.根据权利要求10所述的AON,其中,所述AON包括一个或多个2′-O-甲基硫代膦酸酯寡核苷酸。
12.根据权利要求11所述的AON,包括全长硫代膦酸酯骨架,且其中,所有核苷酸包括2′-O-甲基修饰。
13.根据权利要求1-3中任一项所述的AON,其中,少于10%的核苷酸是核苷酸类似物。
14.一种载体,表达权利要求1至3中任一项所述的AON或其前体。
15.根据权利要求14所述的载体,其为病毒载体。
16.根据权利要求15所述的载体,其为腺相关病毒载体。
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Cited By (3)
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Families Citing this family (66)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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WO2016054615A2 (en) | 2014-10-03 | 2016-04-07 | Cold Spring Harbor Laboratory | Targeted augmentation of nuclear gene output |
GB201503408D0 (en) | 2015-02-27 | 2015-04-15 | Proqr Therapeutics N V | Oligonucleotides |
MA41795A (fr) | 2015-03-18 | 2018-01-23 | Sarepta Therapeutics Inc | Exclusion d'un exon induite par des composés antisens dans la myostatine |
US10370660B2 (en) * | 2015-05-21 | 2019-08-06 | Wings Therapeutics, Inc. | Antisense oligonucleotides to treat Dystrophic Epidermolysis Bullosa |
RS60493B1 (sr) | 2015-09-15 | 2020-08-31 | Nippon Shinyaku Co Ltd | Antismisaona nukleinska kiselina |
MA45819A (fr) | 2015-10-09 | 2018-08-15 | Sarepta Therapeutics Inc | Compositions et méthodes pour traiter la dystrophie musculaire de duchenne et troubles associés |
AU2016334804B2 (en) | 2015-10-09 | 2022-03-31 | University Of Southampton | Modulation of gene expression and screening for deregulated protein expression |
ES2882500T3 (es) | 2015-12-14 | 2021-12-02 | Cold Spring Harbor Laboratory | Oligómeros antisentido para el tratamiento del síndrome de Dravet |
US11096956B2 (en) | 2015-12-14 | 2021-08-24 | Stoke Therapeutics, Inc. | Antisense oligomers and uses thereof |
WO2017136435A1 (en) | 2016-02-01 | 2017-08-10 | The Usa, As Represented By The Secretary, Department Of Health And Human Services Office Of Technology Transfer National Institute Of Health | Compounds for modulating fc-epsilon-ri-beta expression and uses thereof |
MA45328A (fr) | 2016-04-01 | 2019-02-06 | Avidity Biosciences Llc | Compositions acide nucléique-polypeptide et utilisations de celles-ci |
AU2018205259B2 (en) | 2017-01-06 | 2023-09-14 | Avidity Biosciences, Inc. | Nucleic acid-polypeptide compositions and methods of inducing exon skipping |
GB201711809D0 (en) | 2017-07-21 | 2017-09-06 | Governors Of The Univ Of Alberta | Antisense oligonucleotide |
SG11202001103QA (en) * | 2017-08-11 | 2020-03-30 | Agency Science Tech & Res | Method for screening splicing variants or events |
CN111278991B (zh) | 2017-08-25 | 2022-04-01 | 斯托克制药公司 | 用于治疗病况和疾病的反义寡聚体 |
MA51103A (fr) | 2017-12-06 | 2020-10-14 | Avidity Biosciences Inc | Compositions et procédés de traitement de l'atrophie musculaire et de la dystrophie myotonique |
EP3830259A4 (en) | 2018-08-02 | 2022-05-04 | Dyne Therapeutics, Inc. | MUSCLE TARGETING COMPLEXES AND THEIR USES FOR THE TREATMENT OF FACIOSCAPULOHUMERAL MUSCULAR DYSTROPHY |
US11168141B2 (en) | 2018-08-02 | 2021-11-09 | Dyne Therapeutics, Inc. | Muscle targeting complexes and uses thereof for treating dystrophinopathies |
EP3829595A4 (en) | 2018-08-02 | 2022-08-24 | Dyne Therapeutics, Inc. | MUSCLE TARGETING COMPLEXES AND THEIR USES FOR THE TREATMENT OF DYSTROPHINOPATHIES |
CN113286887A (zh) * | 2018-11-02 | 2021-08-20 | 比奥马林技术公司 | 用于肌营养不良蛋白外显子跳跃的双特异性反义低聚核苷酸 |
MX2022007491A (es) | 2019-12-19 | 2022-08-15 | Nippon Shinyaku Co Ltd | Acido nucleico antisentido que permite la omision de exones. |
AU2021237465A1 (en) | 2020-03-19 | 2022-10-13 | Avidity Biosciences, Inc. | Compositions and methods of treating Facioscapulohumeral muscular dystrophy |
TW202208627A (zh) | 2020-05-11 | 2022-03-01 | 美商斯托克治療公司 | 用於病症及疾病之治療的opa1反義寡聚物 |
WO2022173811A1 (en) * | 2021-02-12 | 2022-08-18 | Rosalind Franklin University Of Medicine And Science | Antisense compounds targeting genes associated with cystic fibrosis |
CA3218805A1 (en) | 2021-05-10 | 2022-11-17 | Ziqing QIAN | Compositions and methods for intracellular therapeutics |
WO2022271818A1 (en) | 2021-06-23 | 2022-12-29 | Entrada Therapeutics, Inc. | Antisense compounds and methods for targeting cug repeats |
AU2022298028A1 (en) | 2021-06-23 | 2023-12-21 | National Center Of Neurology And Psychiatry | Combination of antisense oligomers |
US11638761B2 (en) | 2021-07-09 | 2023-05-02 | Dyne Therapeutics, Inc. | Muscle targeting complexes and uses thereof for treating Facioscapulohumeral muscular dystrophy |
US11771776B2 (en) | 2021-07-09 | 2023-10-03 | Dyne Therapeutics, Inc. | Muscle targeting complexes and uses thereof for treating dystrophinopathies |
EP4215614A1 (en) | 2022-01-24 | 2023-07-26 | Dynacure | Combination therapy for dystrophin-related diseases |
Family Cites Families (92)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5034506A (en) | 1985-03-15 | 1991-07-23 | Anti-Gene Development Group | Uncharged morpholino-based polymers having achiral intersubunit linkages |
US5541308A (en) * | 1986-11-24 | 1996-07-30 | Gen-Probe Incorporated | Nucleic acid probes for detection and/or quantitation of non-viral organisms |
US5766847A (en) | 1988-10-11 | 1998-06-16 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Process for analyzing length polymorphisms in DNA regions |
DE3834636A1 (de) | 1988-10-11 | 1990-04-19 | Max Planck Gesellschaft | Verfahren zur analyse von laengenpolymorphismen in dna-bereichen |
US5608046A (en) | 1990-07-27 | 1997-03-04 | Isis Pharmaceuticals, Inc. | Conjugated 4'-desmethyl nucleoside analog compounds |
FR2675803B1 (fr) | 1991-04-25 | 1996-09-06 | Genset Sa | Oligonucleotides fermes, antisens et sens et leurs applications. |
EP0558697A1 (en) | 1991-06-28 | 1993-09-08 | Massachusetts Institute Of Technology | Localized oligonucleotide therapy |
JPH07501204A (ja) | 1991-06-28 | 1995-02-09 | マサチューセッツ インスティテュート オブ テクノロジー | 局所的オリゴヌクレオチド療法 |
US6200747B1 (en) | 1992-01-28 | 2001-03-13 | North Shore University Hospital Research Corp. | Method and kits for detection of fragile X specific, GC-rich DNA sequences |
US5869252A (en) | 1992-03-31 | 1999-02-09 | Abbott Laboratories | Method of multiplex ligase chain reaction |
US5418139A (en) | 1993-02-10 | 1995-05-23 | University Of Iowa Research Foundation | Method for screening for cardiomyopathy |
CA2116280A1 (en) | 1993-03-05 | 1994-09-06 | Marcy E. Macdonald | Huntingtin dna, protein and uses thereof |
US5741645A (en) | 1993-06-29 | 1998-04-21 | Regents Of The University Of Minnesota | Gene sequence for spinocerebellar ataxia type 1 and method for diagnosis |
US5627263A (en) | 1993-11-24 | 1997-05-06 | La Jolla Cancer Research Foundation | Integrin-binding peptides |
DE4342605A1 (de) | 1993-12-14 | 1995-06-22 | Buna Gmbh | Funktionalisierte Olefinhomo- und -copolymere |
US5962332A (en) | 1994-03-17 | 1999-10-05 | University Of Massachusetts | Detection of trinucleotide repeats by in situ hybridization |
WO1995030774A1 (en) | 1994-05-05 | 1995-11-16 | Beckman Instruments, Inc. | Oligonucleotide repeat arrays |
US5968909A (en) | 1995-08-04 | 1999-10-19 | Hybridon, Inc. | Method of modulating gene expression with reduced immunostimulatory response |
US5854223A (en) | 1995-10-06 | 1998-12-29 | The Trustees Of Columbia University In The City Of New York | S-DC28 as an antirestenosis agent after balloon injury |
US6300060B1 (en) | 1995-11-09 | 2001-10-09 | Dana-Farber Cancer Institute, Inc. | Method for predicting the risk of prostate cancer morbidity and mortality |
PL328563A1 (en) | 1996-02-14 | 1999-02-01 | Isis Pharmaceuticals Inc | Oligonucleotides of modified sugar residues |
US6251589B1 (en) | 1996-07-18 | 2001-06-26 | Srl, Inc. | Method for diagnosing spinocerebellar ataxia type 2 and primers therefor |
CA2241173A1 (en) | 1996-10-30 | 1998-05-07 | Srl, Inc. | Cdna fragment of causative gene of spinocerebellar ataxia type 2 |
US5853995A (en) | 1997-01-07 | 1998-12-29 | Research Development Foundation | Large scale genotyping of diseases and a diagnostic test for spinocerebellar ataxia type 6 |
US20020137890A1 (en) | 1997-03-31 | 2002-09-26 | Genentech, Inc. | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
AU7265298A (en) | 1997-04-29 | 1998-11-24 | Trustees Of Boston University | Methods and compositions for targeted dna differential display |
US6329501B1 (en) | 1997-05-29 | 2001-12-11 | Auburn University | Methods and compositions for targeting compounds to muscle |
US6514755B1 (en) | 1998-08-18 | 2003-02-04 | Regents Of The University Of Minnesota | SCA7 gene and methods of use |
US6280938B1 (en) | 1997-08-19 | 2001-08-28 | Regents Of The University Of Minnesota | SCA7 gene and method of use |
US6794499B2 (en) * | 1997-09-12 | 2004-09-21 | Exiqon A/S | Oligonucleotide analogues |
US6130207A (en) | 1997-11-05 | 2000-10-10 | South Alabama Medical Science Foundation | Cell-specific molecule and method for importing DNA into a nucleus |
JP3012923B2 (ja) | 1998-01-26 | 2000-02-28 | 新潟大学長 | Cagリピート病の治療薬 |
KR100280219B1 (ko) | 1998-02-26 | 2001-04-02 | 이수빈 | 삼핵산 반복 서열을 이용한 신경정신 질환의 진단 방법 및 진단 시약 |
US6322978B1 (en) | 1998-04-20 | 2001-11-27 | Joslin Diabetes Center, Inc. | Repeat polymorphism in the frataxin gene and uses therefore |
CA2343934A1 (en) | 1998-09-25 | 2000-04-06 | The Children's Medical Center Corporation | Short peptides which selectively modulate the activity of protein kinases |
US6172216B1 (en) | 1998-10-07 | 2001-01-09 | Isis Pharmaceuticals Inc. | Antisense modulation of BCL-X expression |
US6399575B1 (en) | 1998-11-10 | 2002-06-04 | Auburn University | Methods and compositions for targeting compounds to the central nervous system |
US6133031A (en) | 1999-08-19 | 2000-10-17 | Isis Pharmaceuticals Inc. | Antisense inhibition of focal adhesion kinase expression |
US20030064944A1 (en) * | 2001-06-21 | 2003-04-03 | Isis Pharmaceuticals Inc. | Antisense modulation of transforming growth factor beta receptor II expression |
US20040226056A1 (en) | 1998-12-22 | 2004-11-11 | Myriad Genetics, Incorporated | Compositions and methods for treating neurological disorders and diseases |
US20020049173A1 (en) | 1999-03-26 | 2002-04-25 | Bennett C. Frank | Alteration of cellular behavior by antisense modulation of mRNA processing |
US6379698B1 (en) | 1999-04-06 | 2002-04-30 | Isis Pharmaceuticals, Inc. | Fusogenic lipids and vesicles |
JP2000325085A (ja) | 1999-05-21 | 2000-11-28 | Masafumi Matsuo | デュシェンヌ型筋ジストロフィー治療剤 |
US20030236214A1 (en) | 1999-06-09 | 2003-12-25 | Wolff Jon A. | Charge reversal of polyion complexes and treatment of peripheral occlusive disease |
US6355481B1 (en) | 1999-06-18 | 2002-03-12 | Emory University | Hybridoma cell line and monoclonal antibody for huntingtin protein |
EP1133993A1 (en) | 2000-03-10 | 2001-09-19 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Substances for the treatment of spinal muscular atrophy |
US20020187931A1 (en) | 2000-04-13 | 2002-12-12 | Michael Hayden | Modulating cell survival by modulating huntingtin function |
US6653467B1 (en) | 2000-04-26 | 2003-11-25 | Jcr Pharmaceutical Co., Ltd. | Medicament for treatment of Duchenne muscular dystrophy |
DE60139394D1 (de) | 2000-04-28 | 2009-09-10 | Asklepios Biopharmaceutical In | Für das dystrophin-minigen kodierende dna-sequenzen und verfahren zu deren verwendung |
AU2001259706A1 (en) | 2000-05-09 | 2001-11-20 | Reliable Biopharmaceutical, Inc. | Polymeric compounds useful as prodrugs |
CN1326990A (zh) | 2000-06-07 | 2001-12-19 | 上海博德基因开发有限公司 | 一种新的多肽——人类dna cgg重复结合蛋白16.17和编码这种多肽的多核苷酸 |
US20030124523A1 (en) | 2000-06-22 | 2003-07-03 | Asselbergs Fredericus Alphonsus Maria | Organic compounds |
US6794192B2 (en) | 2000-06-29 | 2004-09-21 | Pfizer Inc. | Target |
RU2165149C1 (ru) | 2000-07-03 | 2001-04-20 | Шапошников Валерий Геннадьевич | Система формования и упаковки изделий из сахарной ваты |
US6727355B2 (en) | 2000-08-25 | 2004-04-27 | Jcr Pharmaceuticals Co., Ltd. | Pharmaceutical composition for treatment of Duchenne muscular dystrophy |
EP1191097A1 (en) | 2000-09-21 | 2002-03-27 | Leids Universitair Medisch Centrum | Induction of exon skipping in eukaryotic cells |
EP1366160B1 (en) | 2000-10-06 | 2008-07-09 | The Regents Of The University Of Michigan | Mini-dystrophin nucleic acid and peptide sequences |
US6623927B1 (en) | 2000-11-08 | 2003-09-23 | Council Of Scientific And Industrial Research | Method of detection of allelic variants of SCA2 gene |
CA2449412C (en) | 2000-11-30 | 2013-04-02 | Uab Research Foundation | Receptor-mediated uptake of peptides that bind the human transferrin receptor |
WO2002092763A2 (en) | 2001-05-11 | 2002-11-21 | Regents Of The University Of Minnesota | Intron associated with myotonic dystrophy type 2 and methods of use |
CA2526831C (en) | 2001-05-18 | 2012-07-31 | Sirna Therapeutics, Inc. | Rna interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (sina) |
US20050277133A1 (en) | 2001-05-18 | 2005-12-15 | Sirna Therapeutics, Inc. | RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA) |
IL143379A (en) | 2001-05-24 | 2013-11-28 | Yissum Res Dev Co | Oligonucleotide against human ache isoform r and its uses |
WO2003013437A2 (en) | 2001-08-07 | 2003-02-20 | University Of Delaware | Compositions and methods for the prevention and treatment of huntington's disease |
DE60230046D1 (de) | 2001-08-10 | 2009-01-08 | Novartis Ag | Peptide, die atherosklerotische schädigungen binden |
US20060074034A1 (en) | 2001-09-17 | 2006-04-06 | Collins Douglas A | Cobalamin mediated delivery of nucleic acids, analogs and derivatives thereof |
KR20030035047A (ko) | 2001-10-29 | 2003-05-09 | (주)바이오코돈 | Bmp-4 유전자 발현을 이용한 편평태선 질환의 치료 및진단방법 |
WO2003037172A2 (en) | 2001-11-01 | 2003-05-08 | Gpc Biotech Inc. | Endothelial-cell binding peptides for diagnosis and therapy |
US20030134790A1 (en) | 2002-01-11 | 2003-07-17 | University Of Medicine And Dentistry Of New Jersey | Bone Morphogenetic Protein-2 And Bone Morphogenetic Protein-4 In The Treatment And Diagnosis Of Cancer |
WO2003069330A1 (en) | 2002-02-11 | 2003-08-21 | The Trustees Of Columbia University In The City Of New York | System and method for identifying proteins involved in force-initiated signal transduction |
US20050096284A1 (en) | 2002-02-20 | 2005-05-05 | Sirna Therapeutics, Inc. | RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA) |
US7771727B2 (en) | 2002-03-01 | 2010-08-10 | The Administrators Of The Tulane Educational Fund | Conjugates of therapeutic or cytotoxic agents and biologically active peptides |
US20040101852A1 (en) | 2002-11-21 | 2004-05-27 | Isis Pharmaceuticals Inc. | Modulation of CGG triplet repeat binding protein 1 expression |
ITRM20020253A1 (it) | 2002-05-08 | 2003-11-10 | Univ Roma | Molecole chimeriche di snrna con sequenze antisenso per le giunzioni di splicing del gene della distrofina e applicazioni terapeutiche. |
EP1380644A1 (en) | 2002-07-08 | 2004-01-14 | Kylix B.V. | The use of specified TCF target genes to identify drugs for the treatment of cancer, in particular colorectal cancer, in which TCF/beta-catenin/WNT signalling plays a central role |
EP1585560A4 (en) | 2002-07-26 | 2011-03-16 | Mirus Bio Corp | ADMINISTRATION OF MOLECULES AND COMPLEXES TO IN VIVO MAMMAL CELLS |
US20050255086A1 (en) | 2002-08-05 | 2005-11-17 | Davidson Beverly L | Nucleic acid silencing of Huntington's Disease gene |
CA2493297A1 (en) | 2002-08-12 | 2004-02-19 | Universite De Sherbrooke | Methods to reprogram splice site selection in pre-messenger rnas |
GB0219143D0 (en) | 2002-08-16 | 2002-09-25 | Univ Leicester | Modified tailed oligonucleotides |
US8090542B2 (en) | 2002-11-14 | 2012-01-03 | Dharmacon Inc. | Functional and hyperfunctional siRNA |
AU2003284638A1 (en) | 2002-11-25 | 2004-06-18 | Nonprofit Organization Translational Research Organization Of Duchenne Muscular Dystrophy | ENA NUCLEIC ACID DRUGS MODIFYING SPLICING IN mRNA PRECURSOR |
GB0228079D0 (en) | 2002-12-02 | 2003-01-08 | Laxdale Ltd | Huntington's Disease |
WO2004083432A1 (en) | 2003-03-21 | 2004-09-30 | Academisch Ziekenhuis Leiden | Modulation of exon recognition in pre-mrna by interfering with the secondary rna structure |
CA2526893C (en) | 2003-05-14 | 2010-10-26 | Japan Science And Technology Agency | Inhibition of the expression of huntingtin gene |
JP5110877B2 (ja) | 2003-06-02 | 2012-12-26 | ワイス・エルエルシー | 神経筋障害を処置するための、コルチコステロイドと組み合わせたミオスタチン(gdf8)インヒビターの使用 |
ES2302898T3 (es) | 2003-07-11 | 2008-08-01 | Lbr Medbiotech B.V. | Transferencia de genes a celulas musculares mediada por el receptor de manosa-6-fosfato. |
JP2007508030A (ja) | 2003-10-14 | 2007-04-05 | カーネル・バイオファーマ・インコーポレイテッド | 血液脳関門を介してpnaを送達するための2相pna結合体 |
US20050191636A1 (en) | 2004-03-01 | 2005-09-01 | Biocept, Inc. | Detection of STRP, such as fragile X syndrome |
WO2005105995A2 (en) | 2004-04-14 | 2005-11-10 | Sirna Therapeutics, Inc. | RNA INTERFERENCE MEDIATED TREATMENT OF POLYGLUTAMINE (POLYQ) REPEAT EXPANSION DISEASES USING SHORT INTERFERING NUCLEIC ACID (siNA) |
AU2005248147A1 (en) | 2004-05-11 | 2005-12-08 | Alphagen Co., Ltd. | Polynucleotides for causing RNA interference and method for inhibiting gene expression using the same |
EP2206781B1 (en) * | 2004-06-28 | 2015-12-02 | The University Of Western Australia | Antisense oligonucleotides for inducing exon skipping and methods of use thereof |
EP1618881A1 (en) | 2004-07-20 | 2006-01-25 | Santhera Pharmaceuticals (Schweiz) GmbH | Use of non-glucocorticoid steroids for the treatment of muscular dystrophy |
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- 2007-05-18 EP EP07729286.0A patent/EP2027267B1/en active Active
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- 2012-03-26 JP JP2012070033A patent/JP2012147790A/ja active Pending
Cited By (4)
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CN107567498A (zh) * | 2015-03-11 | 2018-01-09 | ProQR治疗上市公司Ⅱ | 用于大疱性表皮松解症治疗的与col7a1外显子73配对的寡核苷酸 |
CN107567498B (zh) * | 2015-03-11 | 2021-09-21 | 翼治疗有限公司 | 用于大疱性表皮松解症治疗的与col7a1外显子73配对的寡核苷酸 |
CN112218949A (zh) * | 2018-03-02 | 2021-01-12 | 莱顿大学医学中心附属莱顿教学医院 | 多瘤病毒复制的抑制 |
CN114144524A (zh) * | 2019-04-09 | 2022-03-04 | 恩维萨基因学公司 | 癌症特异性分子及其使用方法 |
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US8361979B2 (en) | 2013-01-29 |
JP2009537129A (ja) | 2009-10-29 |
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CN101448945A (zh) | 2009-06-03 |
NZ572886A (en) | 2011-11-25 |
EP2027267A1 (en) | 2009-02-25 |
EP1857548A1 (en) | 2007-11-21 |
CA2652408A1 (en) | 2007-11-29 |
EP2027267B1 (en) | 2016-11-16 |
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