The Pereira Lab published a STAR Protocols paper

The Pereira Lab published a STAR Protocols method entitled “Protocol for identifying cellular reprogramming minimal networks using combinatorial transcription factor screening”.

Earlier this year, Kurochkin et al. (Cell Systems) presented REPROcode, a single-cell combinatorial transcription factor (TF) screening platform designed to systematically identify and optimize immune cell reprogramming strategies. This platform incorporates an arrayed lentiviral library of 408 barcoded immune TFs, enabling the identification of optimal TF stoichiometry, fidelity enhancers, and regulators of cell fate conversions.

The current protocol describes the step-by-step methodology for performing combinatorial TF screening to resolve TF networks underlying immune cell identity. Altman et al. describe the full experimental workflow, including production of arrayed lentiviral TF libraries, transduction, and reprogramming of human fibroblasts into distinct immune cells. The paper further details the steps to assemble TF combinations, including cell purification, library preparation, sequencing, and downstream analysis. Importantly, this resource emphasizes practical guidance on cell culture, use of the BD Rhapsody system, sequencing considerations, and quality control steps. Clear experimental checkpoints and troubleshooting guidance are included to support users with varying levels of experience. Although demonstrated here for immune cell fate induction, this strategy that can be easily applied to additional starting and target cell types.

First-authored by Abigail Altman, with contributions from Diogo Cabral and Ilia Kurochkin (corresponding author), this protocol merges experimental and computational strategies to advance reprogramming methodologies, providing a foundation for engineering cell fate with precision. Find the full paper here.