US007834.171B2 (12) United States Patent (10) Patent No.: US 7,834,171 B2 Leake et al. (45) Date of Patent: *Nov. 16, 2010 (54) MODIFIED POLYNUCLEOTIDES FOR 5,414,077 A 5/1995 Lin et al. REDUCING OFF-TARGET EFFECTS IN RNA 5.434,257 A 7/1995 Matteucci et al. INTERFERENCE 5,457,191. A 10/1995 Cook et al. 5,457,527 A 10, 1995 Manns (75) Inventors: Devin Leake, Denver, CO (US); Angela 5,459,255 A 10, 1995 Cook et al. R Reynolds, Conifer, CO (US); 5,484,908. A 1/1996 Froehler et al. (USSWANSE IIIlam Varsnail, E.S.CO soulder, 5,489,6775.487,872 A 2/19961/1996 SanghvietHafeman al. 5,502,177 A 3, 1996 Matteucci et al. (73) Assignee: Dharmacon, Inc., Lafayette, CO (US) 5,514,786 A 5/1996 Cook et al. 5,532,130 A 7, 1996 Alu1 (*) Notice: Subject to any disclaimer, the term of this 5,578,718 A 1 1/1996 Cook et al. patent is extended or adjusted under 35 5,580,767 A 12/1996 Cowsert et al. U.S.C. 154(b) by 303 days. 5,587,361. A 12/1996 Cook et al. This patent is Subject to a terminal dis- 5,587.469 A 12/1996 Cook et al. claimer. 5,587,470 A 12/1996 Cook et al. 5,591,721 A 1/1997 Agrawal et al. (21) Appl. No.: 11/857,732 5,594,121 A 1/1997 Froehler et al. 5,596,086 A 1/1997 Matteucci et al. (22) Filed: Sep.19, 2007 5,608,046 A 3/1997 Cook et al. O O 5,610,289 A 3, 1997 Cook et al. (65) Prior Publication Data 5,614,617 A 3, 1997 Cook et al. US 2008/O242851 A1 Oct. 2, 2008 5,635,488 A 6/1997 Cook et al. O O 5,637,573 A 6/1997 Agrawal et al. Related U.S. Application Data 5,644,048 A T/1997 Yalu (63) Continuation of application No. 11/019,831, filed on Dec. 22, 2004, now Pat. No. 7,595,387, which is a continuation-in-part of application No. 10/551.350. filed as application No. PCT/US2004/010343 on Apr. (Continued) 1, 2004, and a continuation of application No. 10/613, 077, filed on Jul. 1, 2003, now abandoned, which is a FOREIGN PATENT DOCUMENTS continuation-in-part of application No. 10/406,908, EP 1 114 623 10, 2001 filed on Apr. 2, 2003, now abandoned. (60) Provisional application No. 60/630,228, filed on Nov. 22, 2004, provisional application No. 60/543,640, filed on Feb. 10, 2004, provisional application No. (Continued) 60,543,661, filed on Feb. 10, 2004, provisional appli- OTHER PUBLICATIONS cation No. 60/542,646, filed on Feb. 6, 2004, provi sional application No. 60/542,668, filed on Feb. 6, European Search Report from European Patent Application No. 2004. 05734330.3, dated Feb. 10, 2009, 4 pages. (51) Int. Cl. (Continued) E. i.e. 3. Primary Examiner Kimberly Chong ( .01) (74) Attorney, Agent, or Firm Kalow & Springut, LLP: (52) U.S. Cl. .................. 536/24.5:536/24.31:536/24.1; Scott D. Locke, Esq. 514/44; 435/6: 435/325; 435/375 (58) Field of Classification Search ....................... None (57) ABSTRACT See application file for complete search history. (56) References Cited Methods and compositions for performing RNA interference U.S. PATENT DOCUMENTS with decreased off-target effects are provided. The methods and compositions permit effective and efficient applications 2. o Shaolnik et al. of RNA interference to applications such as diagnostics and 5. so45 A 8, 1992 Cook et al. therapeutics through the use of modifications to the siRNA. 5. 15 I. 510 A 9, 1992 Stec et al. Uniquely modified siRNAs have been developed that reduce 5. 1 4. 136 A 5, 1993 Lin et al. off-target effects incurred in gene-silencing. 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