CAREER: Enhancing Stem Cell Differentiation with CRISPR Activators via Non-integrating Methods

职业：通过非整合方法利用 CRISPR 激活剂增强干细胞分化

• 批准号: 1943696
• 负责人: Xiaojun Lian
• 金额: $ 50万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1943696&HistoricalAwards=false
• 关键词: CAREER; Enhancing; Stem; Cell; Differentiation

Regenerative medicine uses stem cells to replace damaged tissues, but this approach requires therapeutic cells with known activity. There is currently no method to generate such cells in large quantities. In developing tissues, cell differentiation involves the appearance of growth factors at certain times to orchestrate differentiation. This project proposes to evaluate the use of viruses to activate genes in order to replicate that orchestration. Educational activities and lab tours will be designed to engage undergraduate and graduate students. A ‘‘Stem Cell and Art’’ showcase will highlight artistic, technical and ethical aspects of stem cell research. It will be developed for K-12 students, their families, and the general public.In this project, human induced pluripotent stem cell (iPSC) differentiation will be enhanced. Genes specific to cell lineages will be manipulated epigenetically using dCas9 activators. A Synergistic Activation Mediator (SAM) system will be used to achieve target gene activation in iPSCs. Non-integrating Adeno-associated virus 1 (AAV1) will deliver SAM activators to iPSCs. Genome-scale activation screens to identify novel genes that facilitate endothelial progenitor (EP) differentiation will also be performed. The positive candidates will be used to enhance EP differentiation with AAV1-SAM. The investigators will also use AAV-SAM to promote beta cell differentiation. These results may provide compelling evidence that targeted gene activation using SAM could be employed to enhance iPSC differentiation. This would introduce a novel technology (SAM activators) to the stem cell differentiation toolkit in addition to small molecules and growth factors.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.

再生医学使用干细胞来替代受损组织，但这种方法需要具有已知活性的治疗细胞。 目前还没有大量生产这种细胞的方法。在发育中的组织中，细胞分化涉及生长因子在特定时间的出现以协调分化。该项目建议评估使用病毒激活基因以复制这种编排。教育活动和实验室图尔斯参观将旨在吸引本科生和研究生。“干细胞与艺术”展示将突出干细胞研究的艺术、技术和伦理方面。它将为K-12学生、他们的家庭和公众开发。在这个项目中，人类诱导多能干细胞（iPSC）分化将得到加强。 将使用dCas 9激活剂在表观遗传学上操纵对细胞谱系特异的基因。 协同激活介体（SAM）系统将用于实现iPSC中的靶基因激活。 非整合型腺相关病毒1（AAV 1）将SAM激活剂递送至iPSC。还将进行基因组规模的活化筛选，以鉴定促进内皮祖细胞（EP）分化的新基因。阳性候选物将用于用AAV 1-SAM增强EP分化。 研究人员还将使用AAV-SAM来促进β细胞分化。 这些结果可以提供令人信服的证据，即使用SAM的靶向基因活化可以用于增强iPSC分化。除了小分子和生长因子外，这将为干细胞分化工具包引入一种新技术（SAM激活剂）。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

Engineering chimeric antigen receptor neutrophils from human pluripotent stem cells for targeted cancer immunotherapy.

• DOI: 10.1016/j.celrep.2022.111128
• 发表时间: 2022-07-19
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Chang, Yun;Syahirah, Ramizah;Wang, Xuepeng;Jin, Gyuhyung;Torregrosa-Allen, Sandra;Elzey, Bennett D.;Hummel, Sydney N.;Wang, Tianqi;Li, Can;Lian, Xiaojun;Deng, Qing;Broxmeyer, Hal E.;Bao, Xiaoping
• 通讯作者: Bao, Xiaoping

Robust genome editing via modRNA-based Cas9 or base editor in human pluripotent stem cells.

• DOI: 10.1016/j.crmeth.2022.100290
• 发表时间: 2022-09-19
• 期刊: Cell reports methods

Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems.

• DOI: 10.1016/j.bioactmat.2022.01.046
• 发表时间: 2022-08
• 期刊: Bioactive materials
• 影响因子: 18.9
• 作者: Jiang Y;Hoenisch RC;Chang Y;Bao X;Cameron CE;Lian XL
• 通讯作者: Lian XL