Publications
Protocol for identifying cellular reprogramming minimal networks using combinatorial transcription factor screening
April 28, 2026 / Volume 7, Issue 2
STAR Protocols
Abigail R. Altman, Diogo Pértiga-Cabral, Carlos-Filipe Pereira, Ilia Kurochkin
Related Data:
Combinatorial TF screening profiled by single cell RNA sequencing in Fibroblasts Combinatorial TF screening profiled by single cell RNA sequencing in Cancer CellsResources:
Highlights
- Comprehensive protocol for producing arrayed lentiviral TF libraries
- Guidance for optimizing reprogramming and achieving uniform TF distribution
- Strategy for purifying cells and generating single-cell transcriptomes and barcodes
- Bioinformatics pipeline for detecting barcodes and assembling TF combinations
Summary
Direct reprogramming offers a powerful approach to generate therapeutic cell types, but progress is limited by an incomplete understanding of transcription factor (TF) cooperativity. Here, we present a protocol for performing combinatorial TF screening to resolve reprogramming factor networks that drive cell identity. We describe steps for arrayed lentiviral production, transduction, and reprogramming of human fibroblasts into distinct immune cells. We detail procedures for cell purification, library preparation, sequencing, and analysis to resolve TF combinations and dynamics.
