Academic Journal
Protocol for identifying cellular reprogramming minimal networks using combinatorial transcription factor screening.
| Title: | Protocol for identifying cellular reprogramming minimal networks using combinatorial transcription factor screening. |
|---|---|
| Authors: | Altman AR; Molecular Medicine and Gene Therapy, Science for Life Laboratory, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden; Wallenberg Centre for Molecular Medicine, Lund University, Lund, Sweden., Pértiga-Cabral D; Molecular Medicine and Gene Therapy, Science for Life Laboratory, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden; Wallenberg Centre for Molecular Medicine, Lund University, Lund, Sweden; CNC - Centre for Neuroscience and Cell Biology, University of Coimbra, Largo Marquês Do Pombal, 3004-517 Coimbra, Portugal; CiBB - Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, Coimbra, Portugal; Doctoral Programme in Experimental Biology and Biomedicine, University of Coimbra, Coimbra, Portugal., Pereira CF; Molecular Medicine and Gene Therapy, Science for Life Laboratory, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden; Wallenberg Centre for Molecular Medicine, Lund University, Lund, Sweden; CNC - Centre for Neuroscience and Cell Biology, University of Coimbra, Largo Marquês Do Pombal, 3004-517 Coimbra, Portugal; CiBB - Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, Coimbra, Portugal; Asgard Therapeutics AB, Medicon Village, 223 81 Lund, Sweden., Kurochkin I; Molecular Medicine and Gene Therapy, Science for Life Laboratory, Lund Stem Cell Center, Lund University, BMC A12, 221 84 Lund, Sweden; Wallenberg Centre for Molecular Medicine, Lund University, Lund, Sweden. Electronic address: ilia.kurochkin@med.lu.se. |
| Source: | STAR protocols [STAR Protoc] 2026 Jun 19; Vol. 7 (2), pp. 104527. Date of Electronic Publication: 2026 Apr 28. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: [Cambridge, MA] : Cell Press, [2020]- |
| MeSH Terms: | Transcription Factors*/metabolism , Transcription Factors*/genetics , Cellular Reprogramming*/genetics, Fibroblasts/cytology ; Fibroblasts/metabolism ; Lentivirus/genetics ; Humans |
| Abstract: | 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. For complete details on the use and execution of this protocol, please refer to Kurochkin et al.1. (Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Competing Interests: | Declaration of interests C.-F.P. has equity interests and serves in a management position at Asgard Therapeutics AB, which develops cancer immunotherapies based on DC reprogramming technologies. C.-F.P. is an inventor on granted patents U.S. 11,345,891, JP 7303743, and CN ZL201880005047.3 and on patent application WO2018/185709 held by Asgard Therapeutics that cover the cell reprogramming approach described here. |
| References: | Cell. 2014 Aug 14;158(4):889-902. (PMID: 25126792) Cell Syst. 2026 Jan 21;17(1):101457. (PMID: 41539305) Cell. 2006 Aug 25;126(4):663-76. (PMID: 16904174) Nat Genet. 2016 Mar;48(3):331-5. (PMID: 26780608) Science. 2017 Apr 21;356(6335):. (PMID: 28428369) Sci Immunol. 2018 Dec 7;3(30):. (PMID: 30530727) Science. 2024 Oct 18;386(6719):eadn9083. (PMID: 39236156) Cell Syst. 2020 Nov 18;11(5):509-522.e10. (PMID: 33038298) Nat Commun. 2021 Mar 12;12(1):1659. (PMID: 33712564) Cell. 2023 Jan 5;186(1):209-229.e26. (PMID: 36608654) Nat Methods. 2012 Jul;9(7):671-5. (PMID: 22930834) Mol Syst Biol. 2022 Sep;18(9):e11129. (PMID: 36106915) Nat Genet. 2025 Oct;57(10):2522-2535. (PMID: 41044248) Cell. 2021 Jun 24;184(13):3573-3587.e29. (PMID: 34062119) Ann N Y Acad Sci. 2012 Aug;1266:7-17. (PMID: 22901251) Nat Biotechnol. 2021 Apr;39(4):510-519. (PMID: 33257861) Immunity. 2025 Oct 14;58(10):2419-2438.e13. (PMID: 40885192) Bioinformatics. 2013 Jan 1;29(1):15-21. (PMID: 23104886) |
| Contributed Indexing: | Keywords: Bioinformatics; Flow Cytometry; Immunology; RNAseq; Sequencing; Single Cell; Stem Cells |
| Substance Nomenclature: | 0 (Transcription Factors) |
| Entry Date(s): | Date Created: 20260429 Date Completed: 20260623 Latest Revision: 20260813 |
| Update Code: | 20260813 |
| PubMed Central ID: | PMC13138051 |
| DOI: | 10.1016/j.xpro.2026.104527 |
| PMID: | 42054212 |
| Database: | MEDLINE |
| ISSN: | 2666-1667 |
|---|---|
| DOI: | 10.1016/j.xpro.2026.104527 |