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TcBuster™ Non-Viral Gene Engineering

TcBuster(TM) - A next-generation non-viral gene delivery system

TcBuster 转座子机制概述

 

TcBuster 是一种使得转移并稳定整合目的基因 (GOI) 成为可能的大容量转座子系统。TcBuster 可以替代逆转录病毒载体和慢病毒载体,引起细胞治疗药开发期间时间显著节省。

为何将 TcBuster 用于基因工程?

TcBuster 是一种实现基因稳定转移到哺乳动物细胞中的非病毒转座子系统。它支持快速生成转基因细胞,是概念验证和生产放大工艺开发中的宝贵工具。凭借 TcBuster 进行基因编辑具有广泛的应用,包括:

  • 癌症免疫疗法 – 引入嵌合抗原受体 (CAR) 或 TCR;负向调节分子失活;掩蔽治疗性细胞免遭宿主免疫
  • 基因疗法 – 将基因递送入宿主细胞以纠正遗传缺陷
  • 疾病模型 – 开发携带疾病相关分子变体的动物模型
  • 药物发现 – 调节作为药物靶标的蛋白质表达
  • 诊断工具 – 上调检测传染性疾病和非传染性疾病的蛋白质
  • 疫苗开发 – 稳定过表达病毒蛋白以诱导保护性免疫
Cell Engineering With TcBuster Graphic

TcBuster 与基于病毒的系统比较

使用非病毒方法规避绕开采用慢病毒系统进行基因工程的障碍。TcBuster 的使用避免了常见的病毒法难题,包括转导效率低、试剂可用性不一致和 GMP 级病毒供应链不安全。

基于病毒的基因递送(例如使用慢病毒或 AAV)涉及将 GOI 包装入病毒载体,然后转导靶细胞。可以快速生成TcBuster 转座子并将其电穿孔入靶细胞。如下图所示,TcBuster 实验方案消除了冗长的病毒载体包装步骤。

用 TcBuster 增强您的细胞工程流程以及

  • 缩短您的细胞工程时限并降低延迟风险。
  • 相比基于病毒的标准方法,整合尺寸更大的载货。AAV 通常可以递送 4-5 kb DNA,而 TcBuster 可以递送 >10 kb(足够用于 2 个 TCR 加辅助蛋白)。
  • 维持高效基因转移和稳定表达。这使得快速生成转基因哺乳动物细胞成为可能,次生影响有限且无选择步骤。
  • 降低基因修饰的成本
  • 利用 TcBuster 的灵活性在一次操作中执行额外的基因编辑技术。
timeline of gene engineering with TcBuster compared to virus-based systems

TcBuster™ 非病毒基因工程

从概念验证到工艺开发

Hear From Our Scientists - What Is TcBuster?

观看此视频,聆听我们专家对采用 TcBuster 进行基因工程的介绍。

通过使用 TcBuster 递送您的目的基因,实现基因工程的潜力。基因编辑是在免疫细胞和再生医学疗法中微调细胞表型的强大工具,可以通过在体内提高细胞潜能和持久性来提高治疗有效性。

与我们的专家合作,为您的细胞疗法项目设计 T 细胞、NK 细胞、饲养细胞和诱导性多能干细胞 (iPSC)。

Transfection Efficiency with Non-Viral Gene Delivery

(Left) Transfection efficiency of T cells using TcBuster for CD19 CAR, (Right) Killing efficiency of CAR-T cells generated with TcBuster

Similar CAR expression was achieved in primary human T cells edited with a CD19 CAR using TcBuster or lentivirus as shown by detection with the Fluorokine fluorescent CD19 antigen (left 2 panels). In the right panel, CD19 CAR-T cells engineered using TcBuster (dark blue) or lentivirus (light blue) show similar cytotoxicity against CD19+ Nalm6/luc cells. The non-electroporated controls (gray) do not kill target cells. 

Delivery of large CAR constructs with TcBuster

Transposition Capacity With TcBuster

TcBuster can efficiently deliver both small single CAR constructs and large-sized, multi-cistronic constructs. Activated T cells were transposed with increasing plasmid sizes, expanded for 4 days, and analyzed for CAR expression by flow cytometry. >30% transposition efficiency was achieved with constructs ranging up to 6 kb of insert. 

与我们合作的步骤

Realize the potential of gene engineering efficiently by partnering with our expert team. Whether you are new to the technique or represent a gene engineering core group with limited bandwidth, we will provide the same level of responsive partnership and customer service. We will connect scientist-to-scientist and walk through the entire project with you.

Cell therapy gene engineering with TcBuster gene transfer system

Project scope and experimental design

  • Timelines, milestones, deliverables, and project team
  • Custom requirements for deliverables
  • Timely quote delivery

What to expect throughout the project

  • Accessibility of project manager or technical lead to facilitate discussions
  • Delivery of technical progress reports as defined in the statement of work (SOW)
  • Shipment of cell samples and other materials for your testing

What defines project completion?

  • Shipment of engineered cells in agreed-upon formulation, packaging, and labeling
  • Delivery of characterization reports, documentation, and other deliverables as defined in the SOW
  • Quarantine of extra materials
  • Project offboarding and archiving

Material Transfer Agreements (MTAs)

  • Perform cell engineering with TcBuster in your own lab
  • Research licenses to support long term discovery programs
  • Commercial licenses for both therapeutics and ancillary materials
  • Available to qualified laboratories

Bio-Techne 信仰合作关系的力量。我们与 Fresenius Kabi 和 Wilson Wolf 联袂推出 ScaleReady™,这降低进入免疫细胞疗法领域的障碍,用于所有规格的生产计划及其所有阶段。ScaleReady 通过引起复杂性和成本显著降低,将细胞和基因疗法付诸实践,与此同时一致地提供出众的重复性和细胞质量。

ScaleReady 产品组合支持可扩展的免疫细胞疗法解决方案,为您的免疫细胞项目输送真正的平台、流程和产品连续性。与 Lovo®Cue® 功能闭式体系自动化细胞处理系统匹配时,Bio-Techne 试剂和 G-Rex® 生物反应器使得在小占地面积内高通量平行加工细胞疗法成为可能。

携手 ScaleReady 实现 TcBuster 应用于免疫细胞疗法中。关于 TcBuster 的其他应用,请联系 Bio-Techne 的基因工程专家

Luminary Therapeutics Corporate Logo

Luminary Therapeutics has signed a licence agreement for the use of TcBuster for the manufacturing of gene-modified cell therapies. Luminary Therapeutics will use TcBuster in their LMY-920 program, a BAFF-CAR-T product.

Cell & Gene Therapy article on TcBuster

This article in Cell & Gene Therapy Insights discusses the transposon-based TcBuster platform and its competitive advantages in cell and gene therapy. Directed evolution of hyperactive TcBuster, TcBuster in cell therapies, advantages and applications of TcBuster, Q&A with the authors.

T Cell-Based Therapies eBook

This eBook provides an overview of several biological and manufacturing challenges facing the development and production of T cell therapies. It follows with examples of how Bio-Techne products and services can address these challenges at each stage of the process.

Frequently Asked Questions

请把您的项目告诉我们,我们的一位专家将联系您讨论我们可以如何提供帮助。