Portal’s Universal Delivery Platform Unlocks Novel Workflows

Predictable, Dose-Dependent Uptake via Passive

Mechanoporation technologies achieve robust intracellular delivery while uniquely preserving cellular homeostasis.

Diffusion Preservation of Cellular Homeostasis and Native State

A single-step workflow enabled robust engineering of multiple modalities to achieve simultaneous CRISPR-mediated gene editing and high-magnitude mRNA expression. This multiplexing proficiency provides a powerful framework for complex biological modeling while completely avoiding cargo-specific biases, staggered expression timelines, metabolic burden, and reduced multi-modal yield associated with sequential transfection protocols.

We leveraged this capability to transiently engineer primary human T cells with circRNA constructs encoding a CD19-targeted chimeric antigen receptor (CAR) and membrane-bound interleukin-12 (mbIL-12). This single-step multi-modal delivery yielded rapid onset and high-magnitude co-expression of both therapeutic modalities, leading to enhanced tumor cell killing efficacy while circumventing permanent genomic manipulation and long-term cellular perturbation.

High-Fidelity Multiplexing Without Loss of Stoichiometry

Upon boosting with GFP self-amplifying RNA (saRNA), human iPSCs were used to generate cerebral organoids. Delivery of saRNA via Portal’s platform results in long-lasting GFP positive cells and does not disrupt lineage commitment or structural scaffolding during a 20-day cerebral organoid maturation protocol.

Transient Engineering for Rapid, Complex Cell Therapy

We validated three distinct implementations, serving as a foundation for a myriad of designs that capitalize on live-cell native MOA study: first, delivering the LgBiT protein directly into cells endogenously tagged with HiBiT facilitated precise, real-time kinetic monitoring of PROTAC-mediated target protein degradation; second, non-viral intracellular loading of an otherwise impermeable Src kinase tracer resulted in dose-dependent competitive displacement by Src small-molecule inhibitors; and lastly, intracellular co-delivery of the four antibody components of the Lumit immunoassay enabled live-cell, wash-free detection of intracellular kinase phosphorylation following pervanadate stimulation with signal-to-background ratios comparable to conventional lysate-based Lumit workflow. By eliminating artificial proxy models and preserving native target conformations, post-translational modifications, and cellular pathways, this direct-to-biology framework compresses validation timelines from months to hours while delivering deeper, physiologically accurate insights into candidate drug mechanisms.

Direct-to-Biology Insights for Early-Phase Drug Discovery

The porous 2D architecture of this platform enables full translational utility across operational scales with simple scaling of design footprint to accommodate throughput requirements, providing a unified framework that bridges the gap between early-stage discovery and large-scale therapeutic manufacturing. For high-throughput (HT) discovery applications, the low-pressure requirements of the silicon membrane enable seamless integration with existing automated cell dispensing instrumentation. This approach leverages a distinct attribute of mechanically-mediated poration: membrane closure can take up to a minute or more, allowing for diffusion of cargo into the cell after the poration event, and enabling a workflow in which cells can be mechanoporated during dispensing into a plate which is pre-loaded with cargo. This principle provides the foundation for automated delivery of cargo in 96- and 384-well formats, achieving uniform cargo delivery across thousands of independent samples without the need for specialized high-pressure infrastructure.

Unified Scalability from Research to Clinical Manufacturing

To evaluate the platform’s compatibility with a multi-step clinical manufacturing process, we performed a longitudinal study involving the sequential engineering of primary human T cells over a 10-day expansion period. The workflow commenced with research-scale delivery of CRISPR RNPs for B2M editing in unstimulated T cells, followed by a standard clinical activation protocol, with robust, >80-fold expansion and high viability up to clinical-scale densities. The population then underwent a second, clinical-scale boost of GFP mRNA using a porous membrane integrated in-line with the LOVO system, resulting in high-efficiency editing and near-universal mRNA expression.