The ULTIMATE Reagent: A Universal Photocleavable and Clickable Reagent for the Regiospecific and Reversible End Labeling of Any Nucleic Acid

Jain Piyush K.; Friedman Simon H.

摘要

Abstract >There is a need for methods to chemically incorporate photocleavable labels into synthetic and biologically sourced nucleic acids in a chemically defined and reversible manner. We have previously demonstrated that the light‐cleaved diazo di‐methoxy nitro phenyl ethyl (diazo‐DMNPE) group has a remarkable regiospecificity for modifying terminally phosphorylated siRNA. Building on this observation, we have identified conditions under which a diazo‐DMNPE reagent that we designed (diazo‐DMNPE‐azide or DDA) is able to singly modify any nucleic acid (RNA, DNA, single‐stranded, double‐stranded, 3′ or 5′ phosphate). It can then be modified with any clickable reagent to incorporate arbitrary labels such as fluorophores into the nucleic acid. Finally, native nucleic acid can be regenerated directly through photolysis of the reagent. Use of the described approach should allow for the tagging of any nucleic acid, from any source—natural or unnatural—while allowing for the light‐induced regeneration of native nucleic acid. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>摘要</标题> >需要将光可变标签化学掺入合成和生物源的核酸中的方法一种化学定义和可逆的方式。我们之前已经证明，光切割的DiaZo二甲氧基硝基苯基乙基（Diazo-DMNPE）基团具有显着的终端磷酸化的siRNA的细节性。在这种观察中，我们已经确定了我们设计的Diazo-DMNPE试剂的条件（Diazo-Dmnpe-叠氮化物或DDA）能够单独修饰任何核酸（RNA，DNA，单链，双链，3 '或5'磷酸盐）。然后可以用任何可点击的试剂进行修饰，以将任意标记（例如荧光团）掺入核酸中。最后，可以通过试剂的光解直接再生本地核酸。使用所描述的方法应该允许从任何源自自然或非自然 - 允许任何源自天然或不自然的核酸标记的标记，同时允许天然核酸的光诱导的再生。</ p> </摘要> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-16039/'>《Chembiochem: A European journal of chemical biology》</a> <b style="margin: 0 2px;">|</b><span>2018年第12期</span><b style="margin: 0 2px;">|</b><span>共7页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Jain Piyush K.&option=202" target="_blank" rel="nofollow">Jain Piyush K.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Friedman Simon H.&option=202" target="_blank" rel="nofollow">Friedman Simon H.;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Department of Pharmaceutical SciencesUniversity of Missouri–Kansas City2464 Charlotte Street Kansas City MO 64108 USA;</p> <p>Department of Pharmaceutical SciencesUniversity of Missouri–Kansas City2464 Charlotte Street Kansas City MO 64108 USA;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/179.html" title="分子生物学">分子生物学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=click chemistry&option=203" rel="nofollow">click chemistry;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=DNA&option=203" rel="nofollow">DNA;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=end-labeling&option=203" rel="nofollow">end-labeling;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=nucleic acids&option=203" rel="nofollow">nucleic acids;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=photocleavage&option=203" rel="nofollow">photocleavage;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=RNA&option=203" rel="nofollow">RNA;</a> </p> <div class="translation"> 机译：点击化学;DNA;终点标记;核酸;光学射击;RNA; 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