Improving iPSC reprogramming and CRISPR gene editing workflows and efficacy using CellRaft technology

使用 CellRaft 技术改进 iPSC 重编程和 CRISPR 基因编辑工作流程和功效

• 批准号: 10324993
• 负责人: Jessica Hartman
• 金额: $ 25.66万
• 依托单位: CELL MICROSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324993
• 关键词: Academia; Adherent Culture; Automation; Capital; Case Study; Cell Culture Techniques; Cell Line; Cell Separation; Cell Survival; Cells; Cellular Morphology; Clinical; Clinical Trials; Clone Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Consumption; Custom; DNA Sequence Alteration; Dermal; Development; Disease; Dissociation; Electroporation; Ensure; Equipment; Evaluation; Fibroblasts; Genes; Genetic; Harvest; Human; Image; Industry; Investments; Laboratories; Maintenance; Manuals; Measures; Methodology; Methods; Microscope; Microscopic; Monitor; Morphology; Nature; Pain; Patients; Phase; Phenotype; Plague; Polystyrenes; Population; Process; Protocols documentation; Publishing; Research Personnel; Seminal; Sickle Cell Anemia; Sorting - Cell Movement; Stains; System; Techniques; Technology; Therapeutic; Time; Tissues; Viral; Visualization; base; beta Thalassemia; cell immortalization; cell type; cost; cost effective; cost efficient; culture plates; drug development; drug discovery; feeding; gene therapy; improved; individual patient; induced pluripotent stem cell; instrumentation; microsystems; next generation sequencing; personalized medicine; pluripotency; precision drugs; programs; stem cell growth; stem cells; success; tool

Project Summary Induced pluripotent stem cells (iPSCs) have become a commonly used tool to generate multiple cell types from a given individual patient. Reprogramming non-invasively harvested cells, such as dermal fibroblasts, into iPSCs, allows investigators to generate any cell type from a patient with known genetic and clinical backgrounds. This method has been particularly powerful for cases in which a disease condition manifests in a tissue where acquiring patient-derived primary cells is challenging, or there are genetic mutations. However, the workflows associated with reprogramming, CRISPR gene editing, and differentiating iPSCs are low-throughput, costly, time-consuming, and not supported by commercially available automated instrumentation. Customized automation systems have been published and generally require considerable capital investment, multiple types of equipment, custom programming, and commitment to a single reprogramming method. There is an unmet need in academia and industry for technology that can improve the efficiency and success rates of a) reprogramming, b) iPSC viability, and c) the ability to generate clonally derived CRISPR edited cell lines in an automated fashion. The development of streamlined workflows on a single platform that can deliver higher efficiencies in iPSC reprogramming and cell line development is likely to dramatically enhance the utility and throughput of the technology, thereby accelerating the use of iPSCs in personalized medicine and drug discovery. Cell Microsystems proposes the use of the CellRaft Technology, comprising the automated AIR System and CytoSort Array cell culture consumable as a cost-effective, automated, and gentle solution that can solve many of the pain points that plague these critical workflows. In this Phase I program, we will develop protocols for the culture, reprogramming, and editing of iPSCs using our proprietary consumable and instrumentation and demonstrate that our technology provides faster, more cost effective, and more efficient workflows for iPSC cloning. Commercially, there is a tremendous potential market in laboratories across all industries that perform these workflows, and our company has a proven successful track record in developing tools for the drug discovery market.

项目摘要 诱导性多能干细胞（iPSC）已经成为一种常用的工具， 一个特定的病人。将非侵入性收获的细胞，如真皮成纤维细胞， iPSC允许研究人员从具有已知遗传和临床背景的患者中产生任何细胞类型。 该方法对于其中疾病状况在组织中表现的情况特别有效，其中 获取患者来源的原代细胞具有挑战性，或者存在基因突变。然而，工作流 与重编程、CRISPR基因编辑和分化iPSC相关的基因编辑是低通量的，昂贵的， 耗时，并且不受商业上可获得的自动化仪器的支持。定制 自动化系统已经公开，并且通常需要相当大的资本投资、多种类型 设备，自定义编程，并致力于一个单一的重新编程方法。存在未满足的 学术界和工业界对能够提高效率和成功率的技术的需求a） 重编程的能力，B）iPSC活力，和c）在重组细胞中产生克隆衍生的CRISPR编辑的细胞系的能力。 自动化的方式。在单一平台上开发简化的工作流程， iPSC重编程和细胞系开发的效率可能会大大提高其效用， 该技术的吞吐量，从而加速iPSC在个性化医疗和药物中的使用 的发现Cell Microsystems提出使用CellRaft技术，包括自动化AIR 系统和CytoSort Array细胞培养耗材是一种经济高效、自动化和温和的解决方案， 解决困扰这些关键工作流程的许多痛点。在第一阶段，我们将开发 使用我们专有的耗材和材料， 仪器，并证明我们的技术提供更快，更经济，更有效的 iPSC克隆的工作流程。在商业上，所有实验室都有巨大的潜在市场。 执行这些工作流程的行业，我们公司在开发方面有着成功的记录， 药物发现市场的工具。