Name: single molecule analysis of laser localized psoralen adducts
Uploaded: 2017-04-20T12:30:00
Description: Watch this Scientific Journal Video about Single Molecule Analysis of Laser Localized Psoralen Adducts at JoVE.com

这项研究部分由美国国立卫生研究院，研究所院内研究老龄问题方案（Z01 AG000746-08）和范可尼贫血研究基金的支持。

1.挖-TMP的制备<ol><li> 混合50毫克（0.18毫摩尔）的4&#39;-氯甲基-4,5&#39; ，8-三甲基补骨脂素和590毫克（2.7毫摩尔）-4,7,10-三氧杂-1,13- tridecanedi胺在干燥的25mL圆在氮气下汉字烧瓶中。添加10mL的甲苯，并回流12个小时。在减压下在旋转蒸发器除去溶剂。 <ol><li>纯化在硅胶残余物通过快速柱色谱法。 （1：1：0.5 9），用氯仿，甲醇和28％氨水溶液洗脱该柱。蒸发在减压下在旋转蒸发器除去溶剂...

<ol>
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Cell Biol. 146, 905-916 (1999).</li><li>Lan, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+analysis+of+repair+processes+for+oxidative+DNA+damage+in+mammalian+cells.">In situ analysis of repair processes for oxidative DNA damage in mammalian cells.</a> Proc. Natl. Acad. Sci. U.S.A. 101, 13738-13743 (2004).</li><li>Reynolds, P., Botchway, S. W., Parker, A. W., O'Neill, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+dynamics+of+DNA+repair+proteins+following+laser+microbeam+induced+DNA+damage+-+when+is+a+DSB+not+a+DSB?.">Spatiotemporal dynamics of DNA repair proteins following laser microbeam induced DNA damage - when is a DSB not a DSB?.</a> Mutat. Res. 756, 14-20 (2013).</li><li>Vilenchik, M. M., Knudson, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+DNA+double-strand+breaks:+production,+fidelity+of+repair,+and+induction+of+cancer.">Endogenous DNA double-strand breaks: production, fidelity of repair, and induction of cancer.</a> Proc. Natl. Acad. Sci. U.S.A. 100, 12871-12876 (2003).</li><li>Vilenchik, M. M., Knudson, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation+dose-rate+effects,+endogenous+DNA+damage,+and+signaling+resonance.">Radiation dose-rate effects, endogenous DNA damage, and signaling resonance.</a> Proc. Natl. Acad. Sci. U.S.A. 103, 17874-17879 (2006).</li><li>Aguilera, A., Garcia-Muse, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Causes+of+genome+instability.">Causes of genome instability.</a> Annu. Rev. Genet. 47, 1-32 (2013).</li><li>Swenberg, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+versus+exogenous+DNA+adducts:+their+role+in+carcinogenesis,+epidemiology,+and+risk+assessment.">Endogenous versus exogenous DNA adducts: their role in carcinogenesis, epidemiology, and risk assessment.</a> Toxicol. Sci. 120 (1), S130-S145 (2011).</li><li>Eichman, B. F., Mooers, B. H., Alberti, M., Hearst, J. E., Ho, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+crystal+structures+of+psoralen+cross-linked+DNAs:+drug-dependent+formation+of+holliday+junctions.">The crystal structures of psoralen cross-linked DNAs: drug-dependent formation of holliday junctions.</a> J. Mol. Biol. 308, 15-26 (2001).</li><li>Lai, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+DNA+interstrand+cross-links+and+monoadducts+formed+in+human+cells+induced+by+psoralens+and+UVA+irradiation.">Quantitative analysis of DNA interstrand cross-links and monoadducts formed in human cells induced by psoralens and UVA irradiation.</a> Anal. Chem. 80, 8790-8798 (2008).</li><li>Thazhathveetil, A. K., Liu, S. T., Indig, F. E., Seidman, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psoralen+conjugates+for+visualization+of+genomic+interstrand+cross-links+localized+by+laser+photoactivation.">Psoralen conjugates for visualization of genomic interstrand cross-links localized by laser photoactivation.</a> Bioconjug. Chem. 18, 431-437 (2007).</li><li>Muniandy, P. A., Thapa, D., Thazhathveetil, A. K., Liu, S. T., Seidman, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repair+of+laser-localized+DNA+interstrand+cross-links+in+G1+phase+mammalian+cells.">Repair of laser-localized DNA interstrand cross-links in G1 phase mammalian cells.</a> J Biol. Chem. 284 (41), 27908-27917 (2009).</li><li>Nowsheen, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accumulation+of+oxidatively+induced+clustered+DNA+lesions+in+human+tumor+tissues.">Accumulation of oxidatively induced clustered DNA lesions in human tumor tissues.</a> Mutat. Res. 674, 131-136 (2009).</li><li>Meyer, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clustered+DNA+damage+induces+pan-nuclear+H2AX+phosphorylation+mediated+by+ATM+and+DNA-PK.">Clustered DNA damage induces pan-nuclear H2AX phosphorylation mediated by ATM and DNA-PK.</a> Nucl Acids Res. 41, 6109-6118 (2013).</li><li>Schwab, R. A., Niedzwiedz, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+of+DNA+replication+in+the+vertebrate+model+system+DT40+using+the+DNA+fiber+technique.">Visualization of DNA replication in the vertebrate model system DT40 using the DNA fiber technique.</a> J Vis. Exp. (56), e3255 (2011).</li><li>Kad, N. M., Wang, H., Kennedy, G. G., Warshaw, D. M., Van, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collaborative+dynamic+DNA+scanning+by+nucleotide+excision+repair+proteins+investigated+by+single-+molecule+imaging+of+quantum-dot-labeled+proteins.">Collaborative dynamic DNA scanning by nucleotide excision repair proteins investigated by single- molecule imaging of quantum-dot-labeled proteins.</a> Mol. Cell. 37 (5), 702-713 (2010).</li><li>Spielmann, H. P., Sastry, S. S., Hearst, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+the+large-scale+synthesis+of+psoralen+furan-side+monoadducts+and+diadducts.">Methods for the large-scale synthesis of psoralen furan-side monoadducts and diadducts.</a> Proc. Natl. Acad. Sci. U.S.A. 89 (10), 4514-4518 (1992).</li><li>Huang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+DNA+translocase+FANCM/MHF+promotes+replication+traverse+of+DNA+interstrand+crosslinks.">The DNA translocase FANCM/MHF promotes replication traverse of DNA interstrand crosslinks.</a> Mol. Cell. 52, 434-446 (2013).</li><li>Huang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+Molecule+Analysis+of+Laser+Localized+Interstrand+Crosslinks.">Single Molecule Analysis of Laser Localized Interstrand Crosslinks.</a> Front Genet. 7, 84 (2016).</li><li>Ciccia, A., Elledge, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+DNA+damage+response:+making+it+safe+to+play+with+knives.">The DNA damage response: making it safe to play with knives.</a> Mol. Cell. 40, 179-204 (2010).</li><li>Mori, T., Matsunaga, T., Hirose, T., Nikaido, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+a+monoclonal+antibody+recognizing+ultraviolet+light-induced+(6-4)+photoproducts.">Establishment of a monoclonal antibody recognizing ultraviolet light-induced (6-4) photoproducts.</a> Mutat. Res. 194, 263-270 (1988).</li><li>Poirier, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antisera+specific+for+carcinogen-DNA+adducts+and+carcinogen-modified+DNA:+applications+for+detection+of+xenobiotics+in+biological+samples.">Antisera specific for carcinogen-DNA adducts and carcinogen-modified DNA: applications for detection of xenobiotics in biological samples.</a> Mutat. 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激光定位技术要求与在明亮的视野显微镜可见细胞核使用贴壁细胞。我们已经试图附加非贴壁细胞，如初级淋巴细胞，或松散粘连培养细胞如AD293，与细胞粘附制剂，如聚赖氨酸或胶原，或更复杂的混合物在玻璃表面。虽然这些治疗方法可以将细胞结合到表面，我们发现，他们一般留圆角使其很难集中到细胞核。此外，细胞通常不能存活除去生长培养基后的初始洗涤。然而，对于贴壁细胞的要求?...

DNA是从下外源性试剂如辐射，紫外线，环境毒素，燃烧产物，等。此外，还通过氧化代谢产生的内源性自由基物种攻击恒定攻击。所有这些必须化学或物理破坏DNA的完整性1的潜力。在基因组扰动可以激活DNA损伤反应（DDR），招聘和翻译后修饰级联数百，甚至数千名参与损伤修复蛋白和微RNA，细胞周期，细胞凋亡，衰老和炎症通路的调控2。 大多数的我们对DDR的信息来自于与DNA双链断裂的研究。这在很大程度上是由于技术在活细胞中导入3断裂，包括序列特异性断裂，在基因组DNA的可用性。此外，propensit断裂的y以诱导蛋白质DDR，这可以通过免疫荧光来显示的病灶，已经非常有帮助，用于识别的动力学和响应蛋白质的要求。一个用于研究DDR的关键技术是由邦纳和他的同事，谁在活细胞4的核使用激光束直接DNA双链断裂的条纹在“利益区域”（ROI）推出。实际上，他们创造了一个漫长的焦点，其中DDR的蛋白质可以通过免疫识别。这是由它们的磷酸化组蛋白H2AX的激光暴露的细胞的强条带（γ-H2AX）的示范说明。此后，激光方法已通过DNA双链断裂引起的DDR的众多研究中。虽然功能强大和流行，和戏剧性的免疫荧光图像的来源，应该指出的是，在大多数实验的激光强度进行调整，从而产生可观察到的结果，而对病变身份的关注，密度，或间距。事实上，它可能很难做这类估算。因此，它们在很大程度上忽略了，尽管由激光器5引入DNA损伤的多重性。这有助于在文献6的许多矛盾。 与此相反，以双链断裂，DNA的大多数化学修饰不刺激DDR蛋白质的分离的焦点的形成。这是我们目前的病变频率的理解之光重要的。据估计，在培养的人细胞招致每个细胞周期多达50双链断裂，在S期7，8，9很大程度上形成。形成在非增殖细胞更少。与此相反，核碱基丢失或修改事件，这是在数万每细胞/ 1天，10的数目。因此，我们最大约知道通过比较少见，而那些通过螺旋扭曲病变，合共更为常见诱发少得多的事件引起的DDR。 为了解决有关的细胞反应共价基因组DNA的修改问题，我们希望具有螺旋扭曲的DNA加合物是有内在的DDR诱导活性的工作。此外，为了方便实验设计和解释，我们感兴趣的是其引入可能对于控制时间，是适合于可视化的结构。因此，我们开发了基于补骨脂素的战略。补骨脂素被很好地表征光敏DNA嵌入有利于5&#39; TA：AT网站。不像其他的交联剂如氮芥和丝裂霉素C（MMC）它们不是DNA的反应性，除非暴露于长波紫外光（UVA）光。插层的分子与相反链上，以产生螺旋扭曲间交联胸腺嘧啶碱基（的ICL反应）11。在我们的实验中使用的三甲基补骨脂素大多数产品都是的ICL，相对少的单加成物产生（低于10％）12，以及在一条链上相邻碱基之间的内交联不形成。因为他们是强大的块复制和转录，补骨脂等交联剂，如顺铂和MMC，通常用于化疗。因此补骨脂启用研究，通过一个螺旋扭曲结构跟着DDR的激活，并且还提供了洞察到与临床重要性的化合物的细胞应答。 我们合成，其中三甲基补骨脂素是有联系的地高辛（DIG）的试剂，植物甾醇未在哺乳动物细胞中发现的，并经常用作immunotag。用于光活化的要求在活细胞中的细胞核定位允许通过补骨脂素的ICL在定义的ROI的激光（365纳米）。这些可以通过IMM显示unofluorescence反对挖标签。 DNA修复和DDR蛋白出现在激光的局部的ICL 13,14的条纹。 通过用于产生双链断裂的高激光强度激活DDR可能是由于分离的或聚集损伤15,16。因此，从这些实验结果的相关性，以天然存在的病变，本在低得多的浓度，是不确定的。为了解决有关的DNA加合物补骨脂频率和间距类似的问题，我们利用DNA纤维技术17和immunoquantum点。量子点是比荧光染料更明亮，没有被曝光漂白光。因此，它们经常用于单分子成像18，一个应用程序，其为荧光染料是不够明亮。个人DNA纤维可以上克被拉伸小姑娘滑动，并且可以通过对在细胞收获前的温育过程中掺入的核苷类似物免疫荧光来显示。我们处理的细胞与地高辛补骨脂素和露出的ROI，以激光微照射。纤维从细胞和个体辛补骨脂素加合物准备可以与immunoquantum点进行可视化。将细胞暴露于核苷类似物为相对短的时间（20-60分钟）允许复制大片在激光局部的ICL的附近的显示。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Digoxigenin NHS ester</td>
			<td>Sigma-Aldrich</td>
			<td>11333054001</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloro-psoralen</td>
			<td>Berry and Associates</td>
			<td>PS 5000</td>
			<td></td>
		</tr>
		<tr>
			<td>diaminoglycol</td>
			<td>Sigma-Aldrich</td>
			<td>369519</td>
			<td>4,7,10-Trioxa-1,13-tridecanediamine</td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Acros Organics</td>
			<td>423550040</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Fisher Scientific</td>
			<td>A4524</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium solution</td>
			<td>Sigma-Aldrich</td>
			<td>5002</td>
			<td></td>
		</tr>
		<tr>
			<td>TLC plates</td>
			<td>Analtech, Inc.</td>
			<td>P02511</td>
			<td></td>
		</tr>
		<tr>
			<td>Flass glass column 24/40, 100ml</td>
			<td>Chemglass Life Sciences</td>
			<td>CG-1196-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Nikon T2000_E2 spinning disk confocal microscope, equipped with automated stage and environmental control chamber and plate holder</td>
			<td>Perkin Elmer</td>
			<td></td>
			<td>With Volocity Software</td>
		</tr>
		<tr>
			<td>Micropoint Galvo&#160;</td>
			<td>Andor Technologies</td>
			<td></td>
			<td>with a Nitrogen pulsed laser&#160;</td>
		</tr>
		<tr>
			<td>dye cell</td>
			<td>Andor Technologies</td>
			<td>MP-2250-2-365</td>
			<td></td>
		</tr>
		<tr>
			<td>365 dye</td>
			<td>Andor Technologies</td>
			<td>MP-27-365-DYE</td>
			<td></td>
		</tr>
		<tr>
			<td>IdU&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>17125</td>
			<td></td>
		</tr>
		<tr>
			<td>35mm glass botomm plates 1.5 coverslip, 10mm glass diameter, uncoated</td>
			<td>Matek</td>
			<td>P35G-1.5-10-C</td>
			<td></td>
		</tr>
		<tr>
			<td>microscope slides</td>
			<td>New Comer Supply</td>
			<td>Part # 5070</td>
			<td>New Silane Slides</td>
		</tr>
		<tr>
			<td>Mouse anti BrdU antibody (IdU)</td>
			<td>BD Biosciences</td>
			<td>347580</td>
			<td>1 in 40</td>
		</tr>
		<tr>
			<td>Rat anti BrdU Antibody (CldU)</td>
			<td>Abcam</td>
			<td>ab6326</td>
			<td>1 in 200&#160;</td>
		</tr>
		<tr>
			<td>Rabbit anti Dig antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>710019</td>
			<td>1 in 200</td>
		</tr>
		<tr>
			<td>Q-dot 655 goat anti Rabbit IgG</td>
			<td>ThermoFisher Scientific</td>
			<td>Q-11421MP</td>
			<td>1 in 5000</td>
		</tr>
		<tr>
			<td>AF647- goat anti Rat IgG</td>
			<td>Jackson Immunoresearch</td>
			<td>112-605-167</td>
			<td>1 in 100</td>
		</tr>
		<tr>
			<td>AF488-goat anti mouse IgG</td>
			<td>Jackson Immunoresearch</td>
			<td>115-545-166</td>
			<td>1 in 100</td>
		</tr>
		<tr>
			<td>Zeiss epifluorescent microscope A200</td>
			<td>Zeiss</td>
			<td></td>
			<td>&#160;with Axiovision software</td>
		</tr>
		<tr>
			<td>Q-dot 655 filter</td>
			<td>Chroma</td>
			<td>39107</td>
			<td></td>
		</tr>
	</tbody>
</table>

single molecule analysis of laser localized psoralen adducts

DNA损伤反应（DDR）已经被广泛表征在活细胞中诱导激光微束照射的双链断裂（DSB的）的研究。该DDR到螺旋扭曲共价DNA修饰，包括链间交联的DNA（的ICL），没有作为很好的界定。我们研究通过的ICL刺激DDR，通过immunotagged补骨脂素的光敏激光定位，在活细胞中的细胞核。为了解决约加成分布和复制叉遇到的基本问题，我们结合与其他两种技术的激光定位。 DNA纤维经常用于显示通过在短的脉冲掺入核苷类似物的免疫荧光的复制叉的进展。 Immunoquantum点已被广泛用于单分子成像。在新方法中，从携带激光局部的ICL细胞DNA的纤维铺在显微镜载玻片上。该标记的ICL显示有immunoquantum点和日电子病变间距离来确定。复制叉的碰撞与的ICL可以可视化和不同遭遇的图案识别和定量。

激光局部地高辛TMP（ 图1A）的ICL可以通过对连接到补骨脂该DIG标记的免疫荧光来显示。尽管激光可被引导在任意轮廓的区域撞击，条纹不在细胞“自然”形状，并且合法信号可以从假象由于非特异性结合由一次或二次抗体容易区分。使用不到理想的特异性抗体时，此功能非常有用。的DDR，γ-H2AX的公知的标记物，在挖-TMP的ICL的条纹的一个例子示于图1B。</...

Watch this Scientific Journal Video about Single Molecule Analysis of Laser Localized Psoralen Adducts at JoVE.com

Single Molecule Analysis of Laser Localized Psoralen Adducts

激光器在对DNA损伤的细胞应答的研究经常使用的。然而，它们产生的病变，其间隔，频率，和碰撞与复制叉很少表征。在这里，我们描述了一种方法，使这些参数与激光局部间交联的决心。

激光局部补骨脂素加合物的单分子分析

The DNA Damage Response (DDR) has been extensively characterized in studies of double strand breaks (DSBs) induced by laser micro beam irradiation in live ...

Research

JoVE Journal

Genetics

Laser-assisted Cytoplasmic Microinjection in Livestock Zygotes

激光辅助畜禽受精卵细胞质显微注射

Cytoplasmic microinjection into one-cell embryos is a very powerful technique. As an example, it enables the delivery of genome editing tools that can ...

科研

遗传学

Metagenomic Analysis of Silage

青贮的宏基因组学分析

Metagenomics is defined as the direct analysis of deoxyribonucleic acid (DNA) purified from environmental samples and enables taxonomic identification of ...

Detection of Copy Number Alterations Using Single Cell Sequencing

单细胞测序拷贝数改变的检测

Detection of genomic changes at single cell resolution is important for characterizing genetic heterogeneity and evolution in normal tissues, cancers, and ...

Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen

使用生物素化补骨脂素全球定位RNA-RNA相互作用

Knowing how RNAs interact with themselves and with others is key to understanding RNA based gene regulation in the cell. While examples of RNA-RNA ...

Laser Microirradiation to Study In Vivo Cellular Responses to Simple and Complex DNA Damage

Laser Microirradiation to Study In Vivo Cellular Responses to Simple and Complex DNA Damage

激光 Microirradiation 研究体内细胞对简单复杂 DNA 损伤的反应

DNA damage induces specific signaling and repair responses in the cell, which is critical for protection of genome integrity. Laser microirradiation ...

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

转录因子结合、转录、翻译和营业额的实时分析在细胞激活过程中显示全球事件

Upon activation, cells rapidly change their functional programs and, thereby, their gene expression profile. Massive changes in gene expression occur, for ...

Investigation of Protein Recruitment to DNA Lesions Using 405 Nm Laser Micro-irradiation

405 Nm 激光微辐照对 DNA 损伤的蛋白质招募研究

The DNA Damage Response (DDR) uses a plethora of proteins to detect, signal, and repair DNA lesions. Delineating this response is critical to understand ...

Single Molecule Fluorescence In Situ Hybridization (smFISH) Analysis in Budding Yeast Vegetative Growth and Meiosis

Single Molecule Fluorescence In Situ Hybridization smFISH Analysis in Budding Yeast Vegetative Growth and Meiosis

单分子荧光原位杂交 (smFISH) 在发芽酵母营养生长和减数分裂中的分析

Single molecule fluorescence in situ hybridization (smFISH) is a powerful technique to study gene expression in single cells due to its ability to detect ...

Using In Vitro and In-cell SHAPE to Investigate Small Molecule Induced Pre-mRNA Structural Changes

利用体外和细胞内形状研究小分子诱导的预 mrna 结构变化

In the process of drug development of RNA-targeting small molecules, elucidating the structural changes upon their interactions with target RNA sequences ...