Effect of incorporation of alkyl linkers into siRNAs on RNA interference

Bioorganic and Medicinal Chemistry Letters

Volumn 19, Issue 3, 2009, Pages 875-877

  Ueno, Yoshihito ^a,b   Yoshikawa, Kayo ^a   Kitamura, Yoshiaki ^a   Kitade, Yukio ^a  

^a GIFU UNIVERSITY   (Japan)

^b JAPAN SCIENCE AND TECHNOLOGY AGENCY   (Japan)

Author keywords

Alkyl linker; Off target effect; RNA; RNAi; siRNA

Indexed keywords

ALKYL GROUP; COMPLEMENTARY RNA; SMALL INTERFERING RNA;

ARTICLE; CONTROLLED STUDY; GENE TARGETING; HUMAN; HUMAN CELL; NONHUMAN; RNA CONFORMATION; RNA INTERFERENCE; RNA SEQUENCE; RNA SYNTHESIS; SEQUENCE ANALYSIS;

ANIMALS; CHEMISTRY, PHARMACEUTICAL; CIRCULAR DICHROISM; DRUG DESIGN; GENE EXPRESSION PROFILING; GENE SILENCING; GENOMICS; LUCIFERASES; MODELS, CHEMICAL; OLIGONUCLEOTIDES, ANTISENSE; RENILLA; RNA INTERFERENCE; RNA, SMALL INTERFERING; TEMPERATURE;

EID: 58549119331     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2008.11.107     Document Type: Article

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17. MALDI-TOF/MS analyses of RNAs. Spectra were obtained with a time-of-flight mass spectrometer. ON20: calculated mass, 6703.9; observed mass, 6707.2. ON22: calculated mass, 6731.9; observed mass, 6733.4. ON24: calculated mass, 6759.9; observed mass, 6762.4. ON26: calculated mass, 6426.9; observed mass, 6428.0. ON29: calculated mass, 6853.0; observed mass, 6850.9. ON31: calculated mass, 6881.0; observed mass, 6878.5. ON33: calculated mass, 6909.0; observed mass, 6913.1. ON35: calculated mass, 6536.0; observed mass, 6540.1.
18. 4/mL) were transferred to 96-well plates (100 μL per well). They were transfected, using TransFast (Promega), according to 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 whole 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 cell lysates were determined with a dual-luciferase assay system (Promega) according to a manufacturer's protocol. The results were confirmed by at least three independent transfection experiments with two cultures each and are expressed as the average from four experiments as mean ± SD.