Synthesis of nuclease-resistant siRNAs possessing benzene-phosphate backbones in their 3′-overhang regions

Bioorganic and Medicinal Chemistry Letters

Volumn 18, Issue 19, 2008, Pages 5194-5196

  Ueno, Yoshihito ^a,b   Inoue, Takumi ^a   Yoshida, Mahito ^a   Yoshikawa, Kayo ^a   Shibata, Aya ^a   Kitamura, Yoshiaki ^a   Kitade, Yukio ^a  

^a GIFU UNIVERSITY   (Japan)

^b JAPAN SCIENCE AND TECHNOLOGY AGENCY   (Japan)

Author keywords

Aromatic compound; Nucleobase; RNA; RNAi; siRNA

Indexed keywords

1,3 BIS(HYDROXYMETHYL)BENZENE; BENZENE DERIVATIVE; N 1 [3,5 BIS(HYDROXYMETHYL)PHENYL]THYMINE; NUCLEASE; NUCLEIC ACID BASE; NUCLEOSIDE DERIVATIVE; PHOSPHATE; PHOSPHODIESTERASE; SMALL INTERFERING RNA; SNAKE VENOM; SPLEEN EXONUCLEASE; THYMIDINE; UNCLASSIFIED DRUG;

3' UNTRANSLATED REGION; ARTICLE; DRUG SYNTHESIS; HELA CELL; HUMAN; HUMAN CELL; HYDROLYSIS; IN VITRO STUDY; RNA STRUCTURE;

BASE SEQUENCE; BENZENE DERIVATIVES; ENDORIBONUCLEASES; HELA CELLS; HUMANS; NUCLEIC ACID DENATURATION; PHOSPHORIC ACID ESTERS; PHOSPHORIC DIESTER HYDROLASES; RNA, SMALL INTERFERING; SNAKE VENOMS; STRUCTURE-ACTIVITY RELATIONSHIP; THYMIDINE; TUMOR CELLS, CULTURED; UREA;

EID: 52049089698     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2008.08.086     Document Type: Article

References (17)

1. Fire A., Xu S., Montgomery M.K., Kostas S.A., Driver S.E., and Mello C.C. Nature 391 (1998) 806
2. Elbashir S.M., Harborth J., Lendeckel W., Yalcin A., Weber K., and Tuschl T. Nature 411 (2001) 494
3. Bumcrot D., Manoharan M., Koteliansky V., and Sah D.W.Y. Nat. Chem. Biol. 2 (2006) 711
4. Behlke M.A. Mol. Ther. 13 (2006) 644
5. Mammond S.M., Boettcher S., Caudy A.A., Kobayashi R., and Hannon G.J. Science 293 (2001) 1146
6. Martinez J., Patkaniowska A., Urlaub H., Lührmann R., and Tuschl T. Cell 110 (2002) 563
7. Lingel A., Simon B., Izaurralde E., and Sattler M. Nature 426 (2003) 465
8. Yan K.S., Yan S., Farooq A., Han A., Zeng L., and Zhou M.-M. Nature 426 (2003) 469
9. Song J.-J., Liu J., Tolia N.H., Schneiderman J., Smith S.K., Martienssen R.A., Hannon G.J., and Joshua-Tor L. Nat. Struct. Biol. 12 (2003) 1026
10. Ma J.B., Te K., and Patel D.J. Nature 429 (2004) 318
11. Ueno, Y.; Watanabe, Y.; Shibata, A.; Hirai, M.; Yoshikawa, K.; Takano, T.; Kohara, M.; Kitade, Y., submitted for publication.
12. Ueno Y., Kato T., Sato K., Ito Y., Yoshida M., Inoue T., Shibata A., Ebihara M., and Kitade Y. J. Org. Chem. 70 (2005) 7925
13. Sinha N.D., Biernat J., and Köester H. Tetrahedron Lett. 24 (1983) 5843
14. 15.
15. Puglisi J.D., and Tinoco Jr. I. In: Dahlberg J.E., and Abelson J.N. (Eds). Methods Enzymol. Vol. 180 (1989), Academic press, Inc., San Diego 304-325
16. MALDI-TOF/MS analyses of RNAs. The following spectra were obtained using a time-of-flight mass spectrometer. ON 17: calculated mass, 6571.0; observed mass, 6567.9. ON 18: calculated mass, 6880.3; observed mass, 6877.3.
17. 4/mL) were transferred to 96-well plates (100 μL per well). They were transfected using TransFast (Promega) according to the manufacturer's instructions, for transfection of adherent cell lines. Cells in each well were transfected with a solution (35 μL) of 20 ng of psiCHECK-2 vector (Promega), the indicated amounts of siRNAs, and 0.3 μg of TransFast in Opti-MEM I Reduced-Serum Medium (Invitrogen), and incubated at 37 °C. After 1 h, MEM (100 μL) containing 10% FBS and antibiotics was added to each well, and the mixture was further incubated at 37 °C. After 24 h, cell extracts were prepared in Passive Lysis Buffer (Promega). Activities of firefly and Renilla luciferases in the cell lysates were determined using a dual-luciferase assay system (Promega) according to the manufacturer's protocol. The results were confirmed by performing at least 3 independent transfection experiments with 2 cultures each and are expressed as the average from 4 experiments as mean ± SD.