“實際實驗效果要好於我的預期， ”倪博士說， “MB設計不僅顯著降低了未結合和脫靶探針的螢光， 而且有效減少探針與基因組其它區域的非特異性結合， 後者也非常重要”。
倪博士補充道：“目前的階段性進展雖然突顯了我們在單細胞納米分辨尺度上原位成像人類和小鼠基因組中特異短序列的能力;相信我們團隊建立的方法在接下來研究人類正常和疾病(比如癌症)細胞中3D基因組結構功能中釋放更大的潛力。 ”
張奇偉教授是國際生物資訊學界的權威科學家， 在基因發現、非編碼區及啟動子識別、細胞週期調控、比較基因組學等研究方面有突出成就。
原文連結：
原文摘要：
High-resolution visualization of short non-repetitive DNA in situ in the nuclear genome is essential for studying looping interactions and chromatin organization in single cells. Recent advances in fluorescence in situ hybridization (FISH) using Oligopaint probes have enabled super-resolution imaging of genomic domains with a resolution limit of 4.9 kb. To target shorter elements, we developed a simple FISH method that uses molecular beacon (MB) probes to facilitate the probe-target binding, while minimizing non-specific fluorescence. We used three-dimensional stochastic optical reconstruction microscopy (3D-STORM) with optimized imaging conditions to efficiently distinguish sparsely distributed Alexa-647 from background cellular autofluorescence. Utilizing 3D-STORM and only 29–34 individual MB probes, we observed 3D fine-scale nanostructures of 2.5 kb integrated or endogenous unique DNA in situ in human or mouse genome, respectively. We demonstrated our MB-based FISH method was capable of visualizing the so far shortest non-repetitive genomic sequence in 3D at super-resolution.
作者：張奇偉 點擊：次