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Sequencing of the human genome has opened up several best-kept secrets of the genome. Scientists have identified thousands of genome variations that exist within a population. These variations can be a single nucleotide or a larger chromosomal variation.
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Multiplexed Pan Soluble Ligandome Assaying via OASIS.

Yi-Hung Lee¹, Yesh Doctor¹, Yifan Zhang²

¹Department of Bioengineering, University of California San Diego, CA, USA.

Biorxiv : the Preprint Server for Biology

|January 7, 2026

Summary

We developed Obligate Autocrine Signaling In Situ Screening (OASIS) to enable pooled screening of cell signaling ligands. This method anchors ligands to cells, enforcing autocrine signaling and accelerating ligandome discovery.

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Area of Science:

Cell biology
Molecular biology
Biotechnology

Background:

Screening soluble protein ligands is crucial for understanding cell signaling and drug discovery.
Current methods require arrayed formats due to non-cell-autonomous effects from ligand diffusion.
A need exists for multiplexed pooled assaying of ligands.

Purpose of the Study:

To develop a novel platform, Obligate Autocrine Signaling In Situ Screening (OASIS), for pooled screening of soluble protein ligands.
To enable exclusively autocrine signaling by anchoring ligands to the expressing cell surface.
To accelerate the functional interrogation of the human ligandome.

Main Methods:

Developed OASIS using lentiviral delivery of genetically barcoded ligands fused to a tethering domain.
Anchored proteins to the expressing cell's outer membrane to enforce autocrine signaling.
Validated OASIS with IFNA2 and EGF, and performed a pan-ligandome fitness screen in hiPSCs, followed by single-cell Perturb-Seq.

Main Results:

OASIS demonstrated uncompromised autocrine signaling with significantly reduced paracrine activity.
A pan-ligandome screen identified potent self-renewal factors, including FGF family ligands.
Single-cell Perturb-Seq mapped transcriptional remodeling induced by the ligandome library.

Conclusions:

OASIS is a platform enabling pooled assaying of soluble ligands by enforcing autocrine signaling.
The platform was validated by assaying all human ligands in hiPSCs, measuring fitness and transcriptional impacts.
OASIS facilitates rapid interrogation of ligands and engineered binders for cell signaling studies.