Transcriptional adaptation after deletion of Cdc42 in primary T cells

2025/07/04 by Adam M. Rochussen, Y. Claire, Gillian M. Griffiths · 1 voice
Biochemistry, Genetics and Molecular Biology · Immunology and Microbiology · #CRISPR and Genetic Engineering #Immune Cell Function and Interaction #T-cell and B-cell Immunology

paper · pdf · doi:10.1242/jcs.263826

openalex publication_date 2025/07/04 · openalex created_date 2025/07/05 · openalex updated_date 2026/08/01

Abstract

Cdc42 is a Rho family GTPase known for its central role in cell polarity and cytoskeletal regulation. To understand the role of Cdc42 in polarised secretion from cytotoxic T lymphocytes (CTLs) we used CRISPR/Cas9 gene deletion. Although Cdc42-deleted CTLs initially showed reduced cytotoxicity for up to 2 days after CRISPR-mediated deletion, full secretion and cytotoxicity was rapidly restored and even enhanced while CDC42 protein remained absent. In contrast, chemical inhibition of CDC42 using CASIN consistently decreased secretion in wild-type cells, but had no impact on Cdc42-deleted CTLs, confirming the specificity of this inhibitor. Comparative proteomics and transcriptomics of CTLs after Cdc42 deletion revealed transcriptional changes that could support improved T cell function, including compensation via other Rho GTPases. Targeting the promoter region of Cdc42 did not trigger transcriptional adaptation, consistent with a nonsense-mediated decay mechanism of genetic compensation. Our work highlights the importance of taking orthogonal approaches to study protein function and reveals the remarkable robustness of primary T cells to adapt to loss of an essential gene.

Citations

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Discussions

• If you find a difference in phenotype between knockout and inhibitor/knockdown, transcriptional adaptation may be the culprit. This remarkable phenomenon demonstrates the robustness & complexity of ce [bsky, 7 points, 2 comments]

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