《基於TALE和CRISPR的活細胞染色質DNA螢光標記技術研究》是孫育傑為項目負責人，北京大學為依託單位的面上項目。
基本介紹
- 中文名：基於TALE和CRISPR的活細胞染色質DNA螢光標記技術研究
- 項目類別 ：面上項目
- 項目負責人：孫育傑
- 依託單位 ：北京大學
科研成果,項目摘要,
科研成果
序號 | 標題 | 類型 | 作者 |
|---|---|---|---|
1 | Expansion enhanced nanoscopy | 期刊論文 | Rongqin Li(#); Xuanze Chen; Zixi Lin; Yao Wang; Yujie Sun |
2 | Multicolor Super-resolution Fluorescence Microscopy with Blue and Carmine Small Photoblinking Polymer Dots | 期刊論文 | Chen, Xuanze(#); Liu, Zhihe(#); Li, Rongqin(#); Shan, Chunyan; Zeng, Zhiping; Xue, Boxin; Yuan, Weihong; Mo, Chi; Xi, Peng(*); Wu, Changfeng(*); Sun, Yujie(*) |
3 | GMars-Q Enables Long-Term Live-Cell Parallelized Reversible Saturable Optical Fluorescence Transitions Nanoscopy. | 期刊論文 | Wang, Sheng(#); Chen, Xuanze(#); Chang, Lei(#); Xue, Ruiying; Duan, Haifeng; Sun, Yujie(*) |
4 | Long-term dual-color tracking of genomic loci by modified sgRNAs of the CRISPR/Cas9 system | 期刊論文 | Shao, Shipeng(#); Zhang, Weiwei; Hu, Huan; Xue, Boxin; Qin, Jinshan; Sun, Chaoying; Sun, Yuao; Wei, Wensheng; Sun, Yujie(*) |
5 | Development of bimolecular fluorescence complementation using rsEGFP2 for detection and super-resolution imaging of protein-protein interactions in live cells. | 期刊論文 | Wang, Sheng(#); Ding, Miao(#); Chen, Xuanze; Chang, Lei; Sun, Yujie(*) |
6 | Spying on protein interactions in living cells with reconstituted scarlet light | 期刊論文 | Wang Sheng(#); Ding Miao; Xue Boxin; Hou Yingping; Yujie Sun |
7 | Superior performance with sCMOS over EMCCD in super-resolution optical fluctuation imaging | 期刊論文 | Chen, Xuanze(#); Zeng, Zhiping(#); Li, Rongqin; Xue, Boxin; Xi, Peng(*); Sun, Yujie(*) |
8 | Intra-nucleus Single Molecule Imaging in Living Cells | 期刊論文 | Shipeng Shao(#); Boxin Xue; Yujie Sun |
9 | Live visualization of genomic loci with BiFC-TALE. | 期刊論文 | Hu, Huan(#); Zhang, Hongmin(#); Wang, Sheng(#); Ding, Miao; An, Hui; Hou, Yingping; Yang, Xiaojing; Wei, Wensheng(*); Sun, Yujie(*); Tang, Chao(*) |
10 | Multicolor Photo-Crosslinkable AIEgens toward Compact Nanodots for Subcellular Imaging and STED Nanoscopy | 期刊論文 | Fang, Xiaofeng(#); Chen, Xuanze(#); Li, Rongqin(#); Liu, Zhihe; Chen, Haobin; Sun, Zezhou; Ju, Bo; Liu, Yifei; Zhang, Sean Xiao-An; Ding, Dan; Sun, Yujie(*); Wu, Changfeng(*) |
11 | Illuminating the structure and dynamics of chromatin by fluorescence labeling | 期刊論文 | Shipeng Shao(#); Lei Chang; Yingping Hou; Yujie Sun(*) |
12 | Live Cell Visualization of Multiple Protein-Protein Interactions with BiFC Rainbow | 期刊論文 | Sheng Wang(#); Miao Ding; Boxin Xue; Yingping Hou; Yujie Sun(*) |
13 | Lighting Up Live Cells with Smart Genetically Encoded Fluorescence Probes from GMars Family | 期刊論文 | Wang Sheng(#); Shuai Yao; Sun Chaoying; Xue Boxin; Hou Yingping; Su Xiaodong; Yujie Sun |
14 | Small Photoblinking Semiconductor Polymer Dots for Fluorescence Nanoscopy. | 期刊論文 | Chen, Xuanze(#); Li, Rongqin(#); Liu, Zhihe(#); Sun, Kai; Sun, Zezhou; Chen, Danni; Xu, Gaixia; Xi, Peng; Wu, Changfeng(*); Sun, Yujie(*) |
15 | AgoFISH: cost-effective in situ labelling of genomic loci based on DNA-guided dTtAgo protein | 期刊論文 | Chang, Lei(#); Sheng, Gang; Zhang, Yiwen; Shao, Shipeng; Wang, Yanli(*); Sun, Yujie(*) |
16 | Multiplexed sgRNA Expression Allows Versatile Single Nonrepetitive DNA Labeling and Endogenous Gene Regulation | 期刊論文 | Shipeng Shao(#); Lei Chang; Yuao Sun; Yingping Hou; Xiaoying Fan; Yujie Sun(*) |
17 | GMars-T Enabling Multimodal Subdiffraction Structural and Functional Fluorescence Imaging in Live Cells | 期刊論文 | Wang Sheng(#); Chen Xuanze; Chang Lei; Ding Miao; Xue Ruiying; Duan Haifeng; Sun Yujie |
項目摘要
細胞生命活動中基因位點的動態變化是真核細胞基因表達調控的一個重要層面。因此，特異標記並觀察活細胞基因位點十分重要。傳統的FISH不兼容活細胞，而LacO標記則操作繁瑣通量低，限制了其廣泛套用。TALE和CRISPR的出現為活細胞染色質DNA的標記打開了一扇新的大門。但其目前面臨多個問題，制約了對非重複序列及多基因位點的標記。本項目旨在創新和改進標記和成像方法，利用TALE和CRISPR實現活細胞非重複序列單基因位點的多色標記和成像。在標記方法上，建立基於BiFC的新方法，同時降低背景螢光、脫靶效應以及功能幹擾，並減少所需探針種類降低實驗繁瑣程度。發展sgRNA修飾的方法並與正交CRISPR系統融合實現多色標記。在成像方法上，發展專用於單細胞動態成像的反射式層狀光顯微技術，大幅提信噪比和成像速度，為新標記方法的套用提供技術保證。本項目的完成將為染色質DNA相關動態過程和功能研究提供有力工具。
