EFRI CEE: Optically Controlled Localized Epigenetic Chromatin Remodeling with Photoactivatable CRISPR-dCas9

EFRI CEE：利用光激活 CRISPR-dCas9 进行光控局部表观遗传染色质重塑

• 批准号: 1830878
• 负责人: Lev Perelman
• 金额: $ 200万
• 依托单位: Beth Israel Deaconess Medical Center
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1830878&HistoricalAwards=false
• 关键词: EFRI; CEE; Optically; Controlled; Localized

There are many cancers which come back after apparent recovery and there are others which initially respond to chemotherapy but eventually become treatment-resistant. Along with genetic mutations, this often happens due to epigenetic alterations that do not involve the DNA coding sequence but instead result in a change in gene activity deactivating cellular antitumor systems and activating oncogenes. A recently developed modified form of the gene editing technology called epigenetic CRISPR has the potential to influence those alterations providing effective treatment for recurrent and treatment-resistant cancers.Presently, one of the biggest challenges to using CRISPR that prevents it from being implemented on a systemic level is the concern for off-target effects in normal cells that do not need to be modified. Thus, developing a CRISPR system capable of dynamic control of gene activation in space and time would be a critically important step. To address that, light activated CRISPR was introduced recently. Unfortunately, its major limitation is the need to use blue or UV light, severely reducing the activation penetration depth and, in the case of UV, raising safety concerns. This team address this problem by developing CRISPR-dCas9 which is compatible with NIR light activation, an option not currently available. The proposed light activated chromatin remodeling approach is transformative since it could lead to the development of highly effective targeted epigenetic cancer therapies not prone to off-target activity. In addition, it could also serve as a non-invasive optogenetic tool which does not require a fiber optic implantation and could have various applications in oncology, neurology, ophthalmology, and cardiology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

有许多癌症在明显康复后复发，还有一些癌症最初对化疗有反应，但最终变得耐药。沿着遗传突变，这通常是由于表观遗传改变而发生的，表观遗传改变不涉及DNA编码序列，而是导致基因活性的变化，使细胞抗肿瘤系统失活并激活致癌基因。最近开发的一种名为表观遗传CRISPR的基因编辑技术的改良形式有可能影响这些改变，为复发性和耐药性癌症提供有效治疗。目前，使用CRISPR的最大挑战之一是在不需要修改的正常细胞中的脱靶效应。因此，开发能够在空间和时间上动态控制基因激活的CRISPR系统将是至关重要的一步。为了解决这个问题，最近引入了光激活CRISPR。不幸的是，它的主要限制是需要使用蓝光或UV光，这严重降低了活化渗透深度，并且在UV的情况下，引起了安全问题。该团队通过开发与NIR光激活兼容的CRISPR-dCas 9来解决这个问题，这是一种目前不可用的选择。所提出的光激活染色质重塑方法是变革性的，因为它可以导致开发不倾向于脱靶活性的高效靶向表观遗传癌症疗法。此外，它还可以作为一种非侵入性的光遗传学工具，不需要光纤植入，可以在肿瘤学，神经病学，眼科学和心脏病学中有各种应用。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Two-photon polymerization nanofabrication of ultracompact light scattering spectroscopic probe for detection of pre-cancer in pancreatic cyst

双光子聚合纳米加工超小型光散射光谱探针检测胰腺囊肿癌前病变

• DOI: 10.1016/j.optlaseng.2021.106616
• 发表时间: 2021
• 期刊: Optics and Lasers in Engineering
• 影响因子: 4.6
• 作者: Sheil, Conor J.;Khan, Umar;Zakharov, Yuri N.;Coughlan, Mark F.;Pleskow, Douglas K.;Sawhney, Mandeep S.;Berzin, Tyler M.;Cohen, Jonah M.;Glyavina, Maria;Zhang, Lei
• 通讯作者: Zhang, Lei

Rapid detection and identification of bacteria directly from whole blood with light scattering spectroscopy based biosensor

利用基于光散射光谱的生物传感器直接从全血中快速检测和鉴定细菌

• DOI: 10.1016/j.snb.2021.130489
• 发表时间: 2021
• 期刊: Sensors and Actuators B: Chemical
• 作者: Qiu, Le;Zhang, Lei;Horowitz, Gary L.;Turzhitsky, Vladimir;Coughlan, Mark F.;Glyavina, Maria;Khan, Umar;Zakharov, Yuri N.;Vitkin, Edward;Itzkan, Irving
• 通讯作者: Itzkan, Irving

A portable light scattering system for detecting expelled droplets to assess Covid-19 transmission risk

便携式光散射系统，用于检测排出的液滴以评估 Covid-19 传播风险

• DOI: 10.1117/12.2608902
• 发表时间: 2022
• 期刊: Biomedical Applications of Light Scattering XII
• 作者: Coughlan, Mark F.;Sawhney, Mandeep S.;Pleskow, Douglas K.;Berzin, Tyler M.;Khan, Umar;Glyavina, Maria;Zhang, Xuejun;Chen, Liming;Upputuri, Paul K.;Sheil, Conor J.
• 通讯作者: Sheil, Conor J.

246.8: In Vivo Monitoring of Liver Organoids Differentiation Using Live-Screening-Optical Technologies

246.8：使用实时筛选光学技术体内监测肝脏类器官分化

• DOI: 10.1097/01.tp.0000886176.20263.10
• 发表时间: 2022
• 期刊: Transplantation
• 影响因子: 6.2
• 作者: Pettinato, Giuseppe;Coughlan, Mark F.;Zhang, Xuejun;Chen, Liming;Glyavina, Maria;Zakharov, Yuri N.;Zhang, Lei;Qiu, Le;Fisher, Robert A.;Perelman, Lev T.
• 通讯作者: Perelman, Lev T.