B-Cell Engineering with Portal's Platform

B Cells

Portal powers all of your B cell workflows

CRISPR editing and multiplexed engineering of primary B cells
Transient expression of receptors, transcription factors, and other functional payloads
B cell engineering for antibody discovery, disease modeling, and next-generation cell therapies

Highlighted Data and Applications

Sustained Cell Surface Receptor Expression in B cells Boosted with circular RNA (cRNA)

Compared to ~12% expression in the untreated control, B cells boosted with cRNA of the target surface receptor showed a 4x increase in positivity, reaching ~46% on Day 1 (D1) post-treatment and sustained through at least Day 3 (D3). This result demonstrates that high-efficiency cRNA delivery via Portal’s technology can enable effective target protein expression.

Flow cytometry histograms showing expression of a transduction enhancer RNA in primary human CD19+ B cells after mechanoporation. Control cells (gray) show no fluorescence shift, while Boosted cells (purple) show 73% positive expression, confirming efficient intracellular delivery of the B1 Prep Kit transduction enhancer to B cells.

Comparison of GFP expression (% GFP+ cells) in CD19+ B cells across four conditions. Without lentivirus, both Control and Boost show minimal GFP signal. With B1 lentivirus, Control cells reach ~10% GFP+, while Boost-treated cells achieve ~47% GFP+ — a 3.6-fold improvement demonstrating that mechanoporation-delivered transduction enhancer RNA substantially increases lentiviral gene transfer efficiency in primary B cells.

RNA Delivery Unlocks Efficient B-Cell Engineering

Intracellular delivery of RNA enabled strong functional gains in primary human B cells. Following delivery of a circular RNA encoding a lentiviral transduction enhancer, B cells exhibited robust intracellular expression of the enhancer, with 70%+ of cells positive by flow cytometry. This translated directly into a 3.6-fold improvement in downstream gene transfer: exposure to a GFP-encoding lentivirus resulted in 45%+ GFP+ B cells, up from a ~10% baseline without the enhancer — while no-virus conditions showed minimal background signal. These results demonstrate that RNA pre-treatment can substantially improve lentiviral transduction efficiency in primary B cells without compromising cell integrity.

Flow cytometry histograms showing GFP mRNA expression across four major PBMC subsets after mechanoporation. T cells (CD3+), B cells (CD19+), NK cells (CD56+), and Monocytes (CD14+) each show a clear rightward fluorescence shift in Boosted samples (purple) compared to Control (gray), demonstrating efficient cargo delivery to all subsets — including B cells — directly within mixed, unsorted PBMC populations.

Viability of mixed PBMCs after GFP mRNA delivery. Control cells maintain ~97% viability, while Boosted cells retain ~73% viability (n=4, individual data points shown with error bars), confirming that the delivery process preserves strong cell health across the heterogeneous immune population.

Direct mRNA Expression in B Cells Within Mixed Immune Samples

Efficient RNA delivery was also achieved without prior B-cell isolation. In mixed PBMC populations, delivery of GFP mRNA resulted in a clear increase in GFP expression specifically within the CD19+ B-cell compartment, with minimal signal in untreated controls. This confirms that B cells can be selectively and efficiently programmed directly within heterogeneous immune samples, supporting streamlined workflows and physiologically relevant immune assays.