Efficient sequence-directed psoralen targeting using pseudocomplementary Peptide nucleic acids

TY - JOUR

T1 - Efficient sequence-directed psoralen targeting using pseudocomplementary Peptide nucleic acids

AU - Kim, Ki-Hyun

AU - Fan, Xue-Jun

AU - Nielsen, Peter E.

PY - 2007/1/30

Y1 - 2007/1/30

N2 - A pair of decameric pseudocomplementary PNAs which bind to their mixed purine-pyrimidine sequence target in duplex DNA by double duplex invasion has been synthesized with a derivative of 8-methoxypsoralen conjugated to one of the PNAs. It is shown that this pair of psoralen-conjugated pseudocomplementary PNA oligomers, which target a site in the pBluescriptKS+ vector, upon irradiation with long-wavelength UV light (UVA) with high efficiency and specificity form photoadducts to an adjacent 5'-TA site, and more than 50% of these adducts are DNA interstrand cross-links. Transcription elongation by T7 or T3 RNA polymerase is specifically arrested at the psoralen cross-linking site, yielding more than 90% arrested product. These results emphasize the potential of pseudocomplementary PNA oligomers for highly specific gene targeting, in particular, with respect to sequence-directed psoralen photomodification of double-stranded DNA. Thus, such psoralen-PNA conjugates could be very useful in a range of biology and drug discovery applications.

AB - A pair of decameric pseudocomplementary PNAs which bind to their mixed purine-pyrimidine sequence target in duplex DNA by double duplex invasion has been synthesized with a derivative of 8-methoxypsoralen conjugated to one of the PNAs. It is shown that this pair of psoralen-conjugated pseudocomplementary PNA oligomers, which target a site in the pBluescriptKS+ vector, upon irradiation with long-wavelength UV light (UVA) with high efficiency and specificity form photoadducts to an adjacent 5'-TA site, and more than 50% of these adducts are DNA interstrand cross-links. Transcription elongation by T7 or T3 RNA polymerase is specifically arrested at the psoralen cross-linking site, yielding more than 90% arrested product. These results emphasize the potential of pseudocomplementary PNA oligomers for highly specific gene targeting, in particular, with respect to sequence-directed psoralen photomodification of double-stranded DNA. Thus, such psoralen-PNA conjugates could be very useful in a range of biology and drug discovery applications.

KW - Cross-Linking Reagents/chemistry

KW - DNA/chemistry

KW - DNA-Directed RNA Polymerases

KW - Electrophoretic Mobility Shift Assay

KW - Ficusin/chemistry

KW - Gene Targeting

KW - Nucleic Acid Conformation

KW - Nucleic Acid Heteroduplexes/chemistry

KW - Peptide Nucleic Acids/chemistry

KW - Ultraviolet Rays

U2 - 10.1021/bc0603236

DO - 10.1021/bc0603236

M3 - Journal article

C2 - 17256884

SN - 1043-1802

VL - 18

SP - 567

EP - 572

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 2

ER -