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    COMPOSITIONS AND METHODS FOR INHIBITING GENE SILENCING BY RNA INTERFERENCE
    3.
    发明申请
    COMPOSITIONS AND METHODS FOR INHIBITING GENE SILENCING BY RNA INTERFERENCE 审中-公开
    通过RNA干扰抑制基因沉默的组合物和方法

    公开(公告)号：US20100184209A1

    公开(公告)日：2010-07-22

    申请号：US12279594

    申请日：2007-02-16

    申请人： Annaleen Vermeulen ,  Barbara Robertson ,  Scott Baskerville ,  Christina Yamada ,  Devin Leake ,  Yuriy Fedorov ,  Jon Karpilow ,  Anastasia Khvorova

    发明人： Annaleen Vermeulen ,  Barbara Robertson ,  Scott Baskerville ,  Christina Yamada ,  Devin Leake ,  Yuriy Fedorov ,  Jon Karpilow ,  Anastasia Khvorova

    IPC分类号： C12N5/071 ,  C07H21/02

    CPC分类号： C12N15/111 ,  C12N15/113 ,  C12N2310/11 ,  C12N2310/321 ,  C12N2310/3515 ,  C12N2310/53 ,  C12N2320/50 ,  C12N2310/3521

    摘要： The present invention provides compositions and methods for inhibiting gene silencing by the RNAi pathway. The RNAi inhibitors of the invention have a reverse complement (RC) region to the target molecule of interest (e.g., miRNA) in association with at least one flanking region coupled to either at the 3′ or 5′ end of the RC region. The flanking regions can be single-stranded or can have one or more regions of double stranded nucleic acid with or without a hairpin loop. The RNAi inhibitors described herein can inhibit endogenous targets, including but not limited to microRNAs, or piRNAs, or can be used to inhibit the effects of exogenously introduced molecules, such as synthetic siRNAs, siRNAs expressed from vector constructs (e.g., viral expression systems), or siRNAs generated by enzymatic methods. Inhibition is specific, potent, prolonged, and can be performed on a single target or multiple targets simultaneously.

    摘要翻译： 本发明提供了通过RNAi途径抑制基因沉默的组合物和方法。 本发明的RNAi抑制剂与至少一个与RC区的3'或5'末端连接的侧翼区域具有与目的靶分子（例如miRNA）的反向互补序列（RC）区域。 侧翼区域可以是单链的或可以具有一个或多个具有或不具有发夹环的双链核酸区域。 本文所述的RNAi抑制剂可抑制内源性靶标，包括但不限于微小RNA或piRNA，或可用于抑制外源引入分子如合成siRNAs，由载体构建体（例如病毒表达系统）表达的siRNA的作用， ，或通过酶法产生的siRNA。 抑制是具体的，有效的，延长的，并且可以同时在单个靶或多个靶上进行。

    Modified polynucleotides for use in rna interference
    6.
    发明申请
    Modified polynucleotides for use in rna interference 审中-公开
    用于rna干扰的修饰多核苷酸

    公开(公告)号：US20070167384A1

    公开(公告)日：2007-07-19

    申请号：US10551350

    申请日：2004-04-01

    申请人： Devin Leake ,  Angela Reynolds ,  Anastasia Khvorova ,  William Marshall ,  Yuriy Fedorov ,  Kimberly Nichols

    发明人： Devin Leake ,  Angela Reynolds ,  Anastasia Khvorova ,  William Marshall ,  Yuriy Fedorov ,  Kimberly Nichols

    IPC分类号： A61K48/00 ,  C07J17/00 ,  C07H21/02

    CPC分类号： C12N15/111 ,  C12N15/1137 ,  C12N15/1138 ,  C12N2310/14 ,  C12N2310/315 ,  C12N2310/321 ,  C12N2310/322 ,  C12N2320/11 ,  C12N2330/30 ,  C12Y207/11024 ,  C12Y301/03001 ,  C12N2310/3521

    摘要： Methods and compositions for performing RNA interference comprising a wide variety of stabilized siRNAs suitable for use in serum-containing media and for in vivo applications, such as therapeutic applications, are provided. These siRNAs permit effective and efficient applications of RNA interference to applications such as diagnostics and therapeutics through the use of one or more modifications including orthoesters, terminal conjugates, modified linkages and 2′ modified nucleotides. Uniquely modified siRNAs have been developed that reduces off-target effects incurred in gene-silencing. The modifications include phosphorylation of the first 5′ terminal antisense nucleotide; 2′ carbon modifications of the first and second or first, second, and third 5′ terminal antisense nucleotides; and optionally 2′ carbon modifications of the first and second or first, second, and third 5′ terminal sense nucleotide. Control and exaequo molecules are also provided. siRNA molecules and related control, trackability and exaequo agents with specific stability modifications were developed.

    摘要翻译： 提供了用于进行RNA干扰的方法和组合物，其包含适用于含血清培养基和体内应用（例如治疗应用）的多种稳定的siRNA。 这些siRNA允许通过使用一种或多种修饰，包括原酸酯，末端缀合物，经修饰的连接和2'修饰的核苷酸，有效和有效地应用RNA干扰到诸如诊断和治疗之类的应用。 已经开发了独特修饰的siRNA，其减少在基因沉默中引起的脱靶效应。 修饰包括前5'末端反义核苷酸的磷酸化; 第二个或第二个，第二个和第三个5'末端反义核苷酸的2'碳修饰; 和任选的第二个或第二个，第二个，第二个和第三个5'末端有义核苷酸的2'碳修饰。 还提供对照和exaequo分子。 开发了具有特异性稳定性修饰的siRNA分子和相关对照，可追踪性和exaequo剂。

    MODIFIED POLYNUCLEOTIDES FOR USE IN RNA INTERFERENCE
    7.
    发明申请
    MODIFIED POLYNUCLEOTIDES FOR USE IN RNA INTERFERENCE 审中-公开
    用于RNA干扰的修饰的多核苷酸

    公开(公告)号：US20100197023A1

    公开(公告)日：2010-08-05

    申请号：US12626011

    申请日：2009-11-25

    申请人： Devin Leake ,  Angela Reynolds ,  Anastasia Khvorova ,  William Marshall ,  Yuriy Fedorov ,  Kimberly Nichols

    发明人： Devin Leake ,  Angela Reynolds ,  Anastasia Khvorova ,  William Marshall ,  Yuriy Fedorov ,  Kimberly Nichols

    IPC分类号： C12N15/87 ,  C07H21/04

    CPC分类号： C12N15/111 ,  C12N15/1137 ,  C12N15/1138 ,  C12N2310/14 ,  C12N2310/315 ,  C12N2310/321 ,  C12N2310/322 ,  C12N2320/11 ,  C12N2330/30 ,  C12Y207/11024 ,  C12Y301/03001 ,  C12N2310/3521

    摘要： Methods and compositions for performing RNA interference comprising a wide variety of stabilized siRNAs suitable for use in serum-containing media and for in vivo applications, such as therapeutic applications, are provided. These siRNAs permit effective and efficient applications of RNA interference to applications such as diagnostics and therapeutics through the use of one or more modifications including orthoesters, terminal conjugates, modified linkages and 2′ modified nucleotides. Uniquely modified siRNAs have been developed that reduces off-target effects incurred in gene-silencing. The modifications include phosphorylation of the first 5′ terminal antisense nucleotide; 2′ carbon modifications of the first and second or first, second, and third 5′ terminal antisense nucleotides; and optionally 2′ carbon modifications of the first and second or first, second, and third 5′ terminal sense nucleotide. Control and exaequo molecules are also provided. siRNA molecules and related control, trackability and exaequo agents with specific stability modifications were developed.

    摘要翻译： 提供了用于进行RNA干扰的方法和组合物，其包含适用于含血清培养基和体内应用（例如治疗应用）的多种稳定的siRNA。 这些siRNA允许通过使用一种或多种修饰，包括原酸酯，末端缀合物，经修饰的连接和2'修饰的核苷酸，有效和有效地应用RNA干扰到诸如诊断和治疗之类的应用。 已经开发了独特修饰的siRNA，其减少在基因沉默中引起的脱靶效应。 修饰包括前5'末端反义核苷酸的磷酸化; 第二个或第二个，第二个和第三个5'末端反义核苷酸的2'碳修饰; 和任选的第二个或第二个，第二个，第二个和第三个5'末端有义核苷酸的2'碳修饰。 还提供对照和exaequo分子。 开发了具有特异性稳定性修饰的siRNA分子和相关对照，可追踪性和exaequo剂。

    Identification of toxic nucleotide sequences
    9.
    发明申请
    Identification of toxic nucleotide sequences 审中-公开
    鉴定毒性核苷酸序列

    公开(公告)号：US20050203043A1

    公开(公告)日：2005-09-15

    申请号：US11040553

    申请日：2005-01-21

    申请人： Yuriy Fedorov ,  Jon Karpilow ,  Anastasia Khvorova

    发明人： Yuriy Fedorov ,  Jon Karpilow ,  Anastasia Khvorova

    IPC分类号： C07H21/02 ,  C07H21/04 ,  C12N15/11 ,  C12Q1/68 ,  G01N33/48 ,  G01N33/50 ,  G06F19/00

    CPC分类号： C12Q1/6883 ,  C12N15/111 ,  C12N2310/14 ,  C12N2310/53 ,  C12N2320/10 ,  C12Q2600/142

    摘要： Toxic nucleic acid sequences and methods for identifying, using, and screening libraries for them, including in silico screening, are provided. Compositions of the invention comprise unimolecular and double stranded polynucleotides comprising at least one toxicity region. Toxic sequences of the invention include A/G UUU A/G/U, G/C AAA G/C, and/or GCCA, NUUU, wherein N is any nucleotide, or complements thereof. The invention also provides a method of inducing a toxic response in a cell, comprising introducing into the cell a unimolecular or double stranded polynucleotide comprising at least one toxicity region comprising a sequence selected from the group consisting of A/G UUU A/G/U, G/C AAA G/C, GAAT, and GCCA, NUUU, wherein N is any nucleotide, or a complement of any of the foregoing, wherein said unimolecular or double stranded polynucleotide is at least 5 base pairs long and is comprises a sense and antisense region that are at least substantially complementary.

    摘要翻译： 提供了有毒的核酸序列和用于识别，使用和筛选其文库的方法，包括计算机筛选。 本发明的组合物包含包含至少一个毒性区域的单分子和双链多核苷酸。 本发明的毒性序列包括A / G UUU A / G / U，G / C AAA G / C和/或GCCA，NUUU，其中N是任何核苷酸或其互补序列。 本发明还提供了在细胞中诱导毒性反应的方法，包括向细胞中引入单分子或双链多核苷酸，所述单分子或双链多核苷酸包含至少一个毒性区，其包含选自A / G UUU A / G / U ，G / C AAA G / C，GAAT和GCCA，NUUU，其中N是任何前述的任何核苷酸或补体，其中所述单分子或双链多核苷酸长度为至少5个碱基对并且包含感觉 和至少基本上互补的反义区。

    SNP discriminatory siRNA
    10.
    发明申请
    SNP discriminatory siRNA 审中-公开
    SNP鉴别siRNA

    公开(公告)号：US20050176045A1

    公开(公告)日：2005-08-11

    申请号：US11043637

    申请日：2005-01-25

    申请人： Yuriy Fedorov ,  Angela Reynolds ,  Anastasia Khvorova

    发明人： Yuriy Fedorov ,  Angela Reynolds ,  Anastasia Khvorova

    IPC分类号： C12N15/11 ,  C12N15/113 ,  C12Q1/68 ,  C12N15/85

    CPC分类号： C12N15/1135 ,  C12N15/111 ,  C12N2310/14 ,  C12N2310/321 ,  C12N2310/322 ,  C12N2320/11 ,  C12N2320/51 ,  C12N2310/3521

    摘要： A method of identifying SNP specific siRNA is provided. The method comprises comparing the silencing effect of: (i) at least two SNP containing siRNA in cells that contain a SNP target sequence; (ii) said at least two SNP containing siRNA in cells that contain a wild type target sequence; (iii) at least two non-SNP containing siRNA in cells that contain a SNP target sequence; and (iv) said at least two non-SNP containing siRNA in cells that contain a wild type target sequence. Through the method, SNP specific siRNA can be selected for a diverse set of genes, including the Kras gene.

    摘要翻译： 提供鉴定SNP特异性siRNA的方法。 该方法包括比较以下沉默效应：（i）在含有SNP靶序列的细胞中含有至少两种含有SNP的siRNA; （ii）所述至少两个含有含有野生型靶序列的细胞中含有siRNA的SNP; （iii）在含有SNP靶序列的细胞中至少含有两个含有SNP的siRNA; 和（iv）在含有野生型靶序列的细胞中所述至少两种含有SNP的siRNA。 通过该方法，可以选择SNP特异性siRNA用于多种基因，包括Kras基因。