Carrier PNA for shRNA delivery into cells

Bioorganic and Medicinal Chemistry Letters

Volumn 19, Issue 13, 2009, Pages 3410-3413

  Kitamatsu, Mizuki ^a   Kubo, Takanori ^b   Matsuzaki, Rino ^a   Endoh, Tamaki ^a   Ohtsuki, Takashi ^a   Sisido, Masahiko ^a  

^a OKAYAMA UNIVERSITY   (Japan)

^b YASUDA WOMEN'S UNIVERSITY   (Japan)

Author keywords

Cell penetrating peptide; Peptide nucleic acid; RNA interference

Indexed keywords

CARRIER PEPTIDE NUCLEIC ACID; PEPTIDE NUCLEIC ACID; RNA; SHORT HAIRPIN RNA; UNCLASSIFIED DRUG;

ANIMAL CELL; ARTICLE; CHO CELL; CONTROLLED STUDY; DRUG DELIVERY SYSTEM; GENE EXPRESSION REGULATION; NONHUMAN; PEPTIDE SYNTHESIS; RNA HYBRIDIZATION; RNA INTERFERENCE;

ANIMALS; BASE SEQUENCE; CHO CELLS; CRICETINAE; CRICETULUS; PEPTIDE NUCLEIC ACIDS; RNA INTERFERENCE; RNA, DOUBLE-STRANDED; TRANSITION TEMPERATURE;

EID: 66349113704     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2009.05.031     Document Type: Article

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35. 2 until 40-60% confluence. Before cellular uptake, the cells were incubated at 37 °C for 2 h on a fresh medium containing the carrier PNA/FAM-labeled DNA hybrid. The final concentration of the hybrid in the medium was 10 μM. The cells were then washed three times with PBS and examined under a confocal laser-scanning microscope without fixation.
36. 4 cells/mL), and transferred to 96-well plates (100 μL per well). Transfection of the anti-Luc shRNA in CHO-AA8-Luc cells was carried out by separately adding the carrier PNA (250-500 nM) and then the anti-Luc shRNA (500 nM) into F-12 medium containing 10% FBS. At 2-24 h after the addition of the carrier PNA and the anti-Luc shRNA, cells were lysed by Passive Lysis Buffer (Promega) and divided for subsequent experiments. Firefly luciferase expression in CHO-AA8-Luc cells was measured by FLUOstar OPTIMA (BMG Labtech, Germany) with the addition of a Luciferase Assay Reagent II (Promega). Firefly luciferase expression efficiency in each well was normalized by total protein amount, which was quantified from the remaining lysate by the Protein Assay Kit (BIO-RAD, CA, USA).
37. ® suppressed luciferase expression to 94%.