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Programmable nanobody circuits for cell selection.

Nathan B Wang¹, Albert Blanch-Asensio¹, Hannah Cevasco²

¹Department of Chemical Engineering, MIT, Cambridge, MA 02139, USA.

Biorxiv : the Preprint Server for Biology

|May 13, 2026

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This summary is machine-generated.

We developed a novel protein-based circuit, the Destabilized-nanobody Antigen Selection and Identification Tool (DASIT), for efficient, antigen-specific cell selection. DASIT enables scalable, FACS-free cell enrichment for diverse applications in genome engineering and cell therapy manufacturing.

Area of Science:

Biotechnology
Molecular Biology
Cell Biology

Background:

Efficient isolation of specific cell populations is crucial for advancing genome engineering, pooled screening, and cell therapy manufacturing.
Current methods often face bottlenecks in scalability and specificity, hindering progress in these fields.

Purpose of the Study:

To develop a novel protein-based circuit for antigen-specific cell selection, named DASIT (Destabilized-nanobody Antigen Selection and Identification Tool).
To demonstrate DASIT's capability for scalable, FACS-free cell enrichment across various cell types and applications.
To establish an automation-compatible architecture for complex genetic engineering and cell manufacturing processes.

Main Methods:

Engineered a destabilized nanobody fused to an antibiotic resistance protein, forming the DASIT circuit.

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Utilized antigen binding to stabilize the DASIT circuit, creating a selectable signal for antigen-positive cells.

Applied DASIT for multiplexed, logic-gated integration in human iPSCs, high-throughput CRISPR screening, and phenotypic selection of engineered neurons.

Main Results:

DASIT enables robust selection of antigen-expressing cells across different cell types and antigens.
The system supports both stable integration and transient delivery, allowing recyclable selection without permanent genomic modification.
Demonstrated scalable, FACS-free enrichment in challenging applications, including iPSC engineering, CRISPR screening, and neuronal cell therapy.

Conclusions:

DASIT provides an efficient and scalable solution for antigen-specific cell selection, overcoming key bottlenecks in cell engineering and manufacturing.
Decoupling selection from vector integration offers a flexible and automation-compatible platform for multistep genome engineering and large-scale cell production.
DASIT facilitates advancements in high-throughput screening, cell therapy, and precision genome engineering.