Chemical RNA Editing for Genetic Restoration: The Relationship between the Structure and Deamination Efficiency of Carboxyvinyldeoxyuridine Oligodeoxynucleotides
Chemical RNA Editing for Genetic Restoration: The Relationship between the Structure and Deamination Efficiency of Carboxyvinyldeoxyuridine Oligodeoxynucleotides
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用于遗传恢复的化学 RNA 编辑：羧基乙烯基脱氧尿苷寡脱氧核苷酸的结构与脱氨效率之间的关系
DOI：
10.1111/cbdd.12693
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发表时间：
2016
影响因子：
3
通讯作者：
and Toshifumi Tsukahara
中科院分区：
文献类型：
--
作者：
Vu Thi Luyen;Nguyen Thi Kim Thanh;Md Thoufic Anam Azad;Hitoshi Suzuki;and Toshifumi Tsukahara
含5-羧基乙烯基-2-脱氧尿苷的寡核苷酸（CVU-containing ODNs）通过光交联成功地将胞嘧啶位点特异性转变为尿苷，其具有三个部分：互补序列、发夹环和5-末端光响应性核碱基CVU。使用UV（366 nm）照射进行与含有CVU的ODN的光交联，然后进行热处理以脱氨基。通过光裂解（UV，312 nm）切割交联的核苷酸。使用限制性片段长度多态性和荧光测量分析产物。在以前的研究中，我们已经成功地对合成的单链100-mer ODN靶标（ss 100-nt）和体外合成的全长蓝色荧光蛋白mRNA进行了定点光化学碱基取代。尽管C-to-U位点特异性转换的效率强烈依赖于含CVU的ODNs的序列和结构，但含CVU的ODNs与靶向编辑的脱氨效率之间的关系尚不清楚。因此，在本研究中，我们试图确定含CVU的ODNs的最佳序列和一级结构，用于定点特异性转换。为了评估结构-脱氨效率关系，设计并研究了一系列含八个CVU的ODN。我们发现，最佳的脱氨效率与互补序列长度略大于14 nt和发夹长度为9 nt的ODNs实现。
Oligodeoxynucleotides containing 5‐carboxyvinyl‐2ʹ‐deoxyuridine (CVU‐containing ODNs) for successful site‐specific transition of cytosine to uridine by photo‐cross‐linking have three parts: the complementary sequence, hairpin loop and the 5ʹ‐terminal photoresponsive nucleobaseCVU. Photo‐cross‐linking withCVU‐containing ODNs was performed using UV (366 nm) irradiation, followed by heat treatment for deamination. The cross‐linked nucleotide was cleaved by photosplitting (UV, 312 nm). The products were analyzed using restriction fragment length polymorphism and fluorescence measurements. In previous studies, we have successfully performed site‐directed photochemical base substitution toward a synthetic single‐stranded 100‐mer ODN target (ss100‐nt) andin vitro‐synthesized full‐length blue fluorescent protein mRNA as targets. Although the efficiency of C‐to‐U site‐specific transition strongly depends on the sequence and structure ofCVU‐containing ODNs, the relationship betweenCVU‐containing ODNs and the deamination efficiency of targeted editing remains unclear. Therefore, in this study, we attempted to identify the optimal sequence and primary structure ofCVU‐containing ODNs for site‐directed specific transition. To evaluate the structure–deamination efficiency relationship, a series of eightCVU‐containing ODNs were designed and studied. We showed that the optimal deamination efficiency was achieved with ODNs having a complementary sequence length slightly more than 14 nt and a hairpin length of 9 nt.
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影响因子：
16.6
作者：
Nishikura K
通讯作者：
Nishikura K
Masayuki Hayashi、Isao Saito：通过可逆 DNA 光连接将胞嘧啶位点特异性转变为尿嘧啶。
DOI：
--
发表时间：
2006
期刊：
Chemical Communications 30
影响因子：
--
作者：
Kenzo;Fujimoto;Shigeo;Matsuda;Yoshinaga;Yoshimura;Takashi;Matsumura
通讯作者：
Matsumura
影响因子：
16
作者：
Aphasizheva I;Maslov D;Wang X;Huang L;Aphasizhev R
通讯作者：
Aphasizhev R
使用化学 RNA 编辑将蓝色荧光蛋白变为绿色荧光蛋白作为遗传恢复的新策略
影响因子：
3
作者：
Luyen T. Vu;Thanh T. K. Nguyen;Shafiul Alam;Takashi Sakamoto;Kenzo Fujimoto;Hitoshi Suzuki and Toshifumi Tsukahara
通讯作者：
Hitoshi Suzuki and Toshifumi Tsukahara