Allogeneic MicroRNA Editing Cell Development Service

Unlocking Allogeneic CAR-T Potential Through MicroRNA Engineering

The development of allogeneic CAR-T cell therapies offers transformative promise for treating hematological malignancies and solid tumors. A major barrier in this field is balancing immune compatibility and therapeutic potency. To address this, microRNA editing has emerged as a highly efficient and multiplexed gene regulation strategy that enables precise modulation of T cell functionality. Our proprietary miCAR-T technology enables the silencing of multiple endogenous genes while introducing CAR constructs into donor-derived T cells. This approach enhances resistance to the immunosuppressive tumor microenvironment, mitigates graft-versus-host risks, and increases in vivo persistence—critical factors for long-term therapeutic success.

Fig.1 Schematic diagram of miRNA editing.¹

Our Service

We provide end-to-end microRNA editing services tailored for allogeneic CAR-T development, integrating our proprietary Compact Multi-miRNA Module (C3M) platform. Our core offerings include:

• Design & Construction of Custom miCAR-T Vectors

Incorporation of multiple hairpin-based miRNA sequences to enable simultaneous silencing of up to six target genes (e.g., PD-1, CTLA-4, TGFβR, HLA class I/II).

• Donor T Cell Isolation & Activation

High-purity T cell extraction from healthy donors, followed by activation using anti-CD3/CD28 beads.

• Lentiviral or Electroporation-Based Transduction

Introduction of CAR constructs and C3M modules using optimized delivery protocols for high efficiency and minimal off-target effects.

• In Vitro Validation & Characterization

Assessment of gene knockdown efficiency, CAR expression levels, cytokine profiles, cytotoxicity assays, and immunophenotyping.

• Preclinical Evaluation Models

In vivo tumor challenge studies in xenograft models to evaluate persistence, safety, and efficacy of engineered miCAR-T cells.

Why Choose Us

• Multiplexed Gene Silencing

Simultaneous suppression of multiple immune checkpoints or allo-reactivity-associated genes using a single C3M cassette.

• Compact Vector Footprint

The C3M system occupies minimal vector space, allowing co-expression with CARs and other payloads without compromising transduction efficiency.

• Enhanced Tumor Microenvironment Resistance

Knockdown of immunosuppressive signaling pathways enables improved survival and activity in hostile tumor niches.

• Reduced Risk of Host Rejection

Silencing of MHC molecules and co-stimulatory ligands significantly lowers allo-immunogenicity and host-versus-graft responses.

• Scalability for Off-the-Shelf Use

Process-compatible with large-scale GMP production and cryopreservation, enabling robust clinical-grade allogeneic cell therapy pipelines.

Workflow

To streamline your research process, we've designed a transparent and customizable workflow:

Published Data

Several peer-reviewed publications and conference abstracts have validated the concept of microRNA multiplexing for CAR-T cell engineering. Our C3M system draws from optimized structures shown to enhance knockdown stability and minimize off-target expression. In preclinical studies, miCAR-T cells engineered with three or more miRNAs exhibited:

• Up to 95% gene silencing efficiency for individual targets
• Superior tumor clearance in immunocompromised xenograft models
• Reduced expression of exhaustion markers such as PD-1 and TIM-3
• Longer in vivo persistence, linked to memory-like phenotypic shifts

Contact us to request custom validation using your gene targets.

Frequently Asked Questions

Q1: How many genes can be silenced simultaneously using the C3M system?

A1: Up to six gene targets can be silenced concurrently using a single C3M construct, depending on promoter strength and vector design.

Q2: Can the miCAR-T approach be applied to solid tumors?

A2: Yes. We have experience customizing gene silencing modules to overcome solid tumor immunosuppression and T cell exclusion mechanisms.

Contact Us

At Creative Biolabs, we are committed to advancing innovative tools for next-generation immunotherapy. Whether you're developing an early-stage CAR-T proof-of-concept or scaling for IND-enabling studies, our team is ready to support you with precision, speed, and regulatory know-how.

Reference

1. Correia de Sousa, Marta et al. "Deciphering miRNAs' Action through miRNA Editing." International journal of molecular sciences vol. 20,24 6249. Distributed under Open Access License CC BY 4.0, without modification.