Photodynamic antisense regulation of mRNA having a point mutation with psoralen-conjugated oligonucleotide.
Nucleic acid-based drugs, such as antisense oligonucleotide, ribozyme, and small interfering RNA, are specific compounds that inhibit gene expression at the post-transcriptional level. To develop more effective nucleic acid-based drugs, we focused on photo-reactive antisense oligonucleotides. We have optimized the structure of psoralen-conjugated oligonucleotide to improve their sequence selectivity and photo-crosslinking efficiency. Previously, we reported that photo reactive oligonucleotides containing 2'-O-psoralenyl-methoxyethyl adenosine (2'-Ps-eom) showed drastic photo-reactivity with a strictly sequence specific manner in vitro. In this report, we evaluated the binding ability toward intracellular target mRNA. The 2'-Ps-eom selectively photo-cross-linked to the target mRNA extracted from cells. The 2'-Ps-eom also cross-linked to target mRNA in cells. Furthermore, 2'-Ps-eom did not cross-link to mRNA having a mismatch base. These results suggest that 2'-Ps-eom is a powerful antisense molecule to inhibit the expression of mRNA having a point mutation.
Higuchi
Maiko
M
Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, Japan.
Yamayoshi
Asako
A
Kobori
Akio
A
Murakami
Akira
A
D000241:Adenosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000528:radiation effects; D000818:Animals; D011564:Furocoumarins / Q000737:chemistry* / Q000528:radiation effects; D005786:Gene Expression Regulation; D051379:Mice; D041681:NIH 3T3 Cells; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*; D017354:Point Mutation; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D014466:Ultraviolet Rays
C522099:2'-(4,5',8-trimethylpsoralen-4-yl)adenosine; D011564:Furocoumarins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000241:Adenosine