IR@PKUHSC  > 北京大学第一临床医学院  > 核医学科
学科主题临床医学
Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA
Kang, Lei1; Xu, Xiao-jie2; Ma, Chao1; Wang, Rong-fu1; Yan, Ping1; Zhang, Chun-li1; Sun, Hong-wei1; Li, Ding3
关键词Technium-99m MicroRNA Radiolabel Anti-miRNA oligonucleotide (AMO)
刊名CELL BIOCHEMISTRY AND BIOPHYSICS
2015-03-01
DOI10.1007/s12013-014-0281-1
71期:2页:905-912
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemistry & Molecular Biology ; Biophysics ; Cell Biology
研究领域[WOS]Biochemistry & Molecular Biology ; Biophysics ; Cell Biology
关键词[WOS]IN-VIVO ; TC-99M ; TARGETS ; RNA ; RADIOPHARMACEUTICALS ; OLIGONUCLEOTIDES ; OLIGOMERS ; DELIVERY ; TUMORS
英文摘要

Antisense oligonucleotides have been used for a variety of purposes in microRNA (miRNA) research including functional evaluation, target recognition, and gene studies. Although several Tc-99m-radiolabeled oligonucleotides have been reported in antisense imaging, none of those were related to miRNA tracing. Moreover, separation after labeling was always required to achieve acceptable radiochemical purity. In this study, we prepared a Tc-99m-radiolabeled oligonucleotide under optimized conditions for the purpose of tracing miRNA. A 22mer anti-miRNA oligonucleotide (AMO) was designed completely complementary to the sequence of mature miR-21. AMO probe modified with 2′-O-Methyl and phosphorothioate backbone was designed and synthesized. This probe was conjugated with a bifunctional chelator S-acetylmercaptoacetyltriglycine (NHS-MAG3) via a primary amine on 5′-end. Furthermore, it was radiolabeled and its optimization labeling conditions were performed by varying the amount of stannous ion, Tc-99m-pertecnetate, and reaction time, respectively. Finally, the labeled product was identified by gel electrophoresis and evaluated for its serum stability. The AMO was synthesized with partial 2′-OMe and phosphorothioate modification to improve its stability. Excess of MAG3 impurity was removed by precipitation of tin and MAG3 after the conjugation. The labeling efficiency reached 97 % under the optimal reaction conditions of 2 mu g/mu L SnCl2 center dot 2H(2)O addition, Tc-99m solution with high specific activity, and 90-min reaction at room temperature. Gel electrophoresis confirmed that the peak of radioactivity located the same position of oligomer, which identified the successful radiolabeling. After incubated with human fresh serum for 12 h, labeled AMO showed good stability with high radiochemical purity and no significant degradation. A Tc-99m-labeled AMO targeting miR-21 can be prepared with high labeling efficiency under optimized conditions, which provides a good support for the future use of miRNA-targeted tracing and imaging.

语种英语
WOS记录号WOS:000349241400050
项目编号NSFC 81101065 ; 31100604 ; 81441051 ; 81472589 ; 2011YQ03011409 ; 20110001120043 ; Z141107002514159 ; Z141102001814055
资助机构National Natural Science Foundation of China ; National Major Scientific Equipment Special Fund ; Higher Education Doctoral Program of China ; Beijing Capital Special Development Application Program ; Beijing Nova Program
引用统计
被引频次:2[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/65950
专题北京大学第一临床医学院_核医学科
作者单位1.Peking Univ, Hosp 1, Dept Nucl Med, Beijing 100034, Peoples R China
2.Beijing Inst Biotechnol, Dept Med Mol Biol, Beijing 100850, Peoples R China
3.261 Hosp PLA, Ctr Biol Diag & Therapy, Beijing 100094, Peoples R China
推荐引用方式
GB/T 7714
Kang, Lei,Xu, Xiao-jie,Ma, Chao,et al. Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA[J]. CELL BIOCHEMISTRY AND BIOPHYSICS,2015,71(2):905-912.
APA Kang, Lei.,Xu, Xiao-jie.,Ma, Chao.,Wang, Rong-fu.,Yan, Ping.,...&Li, Ding.(2015).Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA.CELL BIOCHEMISTRY AND BIOPHYSICS,71(2),905-912.
MLA Kang, Lei,et al."Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA".CELL BIOCHEMISTRY AND BIOPHYSICS 71.2(2015):905-912.
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[Kang, Lei]的文章
[Xu, Xiao-jie]的文章
[Ma, Chao]的文章
百度学术
百度学术中相似的文章
[Kang, Lei]的文章
[Xu, Xiao-jie]的文章
[Ma, Chao]的文章
必应学术
必应学术中相似的文章
[Kang, Lei]的文章
[Xu, Xiao-jie]的文章
[Ma, Chao]的文章
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。