Photomodulating RNA cleavage using photolabile circular antisense oligodeoxynucleotides.
Caged antisense oligodeoxynucleotides (asODNs) are synthesized by linking two ends of linear oligodeoxynucleotides using a photocleavable linker. Two of them (H30 and H40) have hairpin-like structures which show a large difference in thermal stability (Delta T(m) = 17.5 degrees C and 11.6 degrees C) comparing to uncaged ones. The other three (C20, C30 and C40) without stable secondary structures have the middle 20 deoxynucleotides complementary to 40-mer RNA. All caged asODNs have restricted opening which provides control over RNA/asODN interaction. RNase H assay results showed that 40-mer RNA digestion could be photo-modulated 2- to 3-fold upon light-activation with H30, H40, C30 and C40, while with C20, RNA digestion was almost not detectable; however, photo-activation triggered >20-fold increase of RNA digestion. And gel shift assays showed that it needed >0.04 microM H40 and 0.5 microM H30 to completely bind 0.02 microM 40-mer RNA, and for C40 and C30, it needed >0.2 microM and 0.5 microM for 0.02 microM 40-mer RNA binding. However, even 4 microM C20 was not able to fully bind the same concentration of 40-mer RNA. By simple adjustment of ring size of caged asODNs, we could successfully photoregulate their hybridization with mRNA and target RNA hydrolysis by RNase H with light activation.
Tang
XinJing
X
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University No. 38, Xueyuan Rd, Beijing 100191, China. xinjingt@bjmu.edu.cn
Su
Meng
M
Yu
LiLi
L
Lv
Cong
C
Wang
Jie
J
Li
ZhongJin
Z
D004270:DNA, Circular / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry* / Q000528:radiation effects; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism; D014466:Ultraviolet Rays
D004270:DNA, Circular; D020319:Oligodeoxyribonucleotides, Antisense; D012313:RNA; D016914:Ribonuclease H