Cheong, Sek-ShirSek-ShirCheongLuis, TiagoTiagoLuisStewart, MichelleMichelleStewartHillier, RosieRosieHillierHind, MatthewMatthewHindDean, CharlotteCharlotteDean2023-11-012023-11-012023-11Disease Models and Mechanisms, 2023, 16 (11)1754-8403http://hdl.handle.net/10044/1/107301Precision-Cut Lung Slices (PCLS) are used for a variety of applications. However, methods to manipulate genes in PCLS are currently limited. We developed a novel method, TAT-Cre Recombinase-mediated floxed Allele modification in Tissue Slices (TReATS), to induce highly effective and temporally controlled gene deletion or activation in ex vivo PCLS. Treatment of PCLS from Rosa26-flox-stop-flox-EYFP mice with cell-permeant TAT-Cre recombinase induced ubiquitous EYFP protein expression, indicating successful Cre-mediated excision of the upstream loxP-flanked stop sequence. Quantitative real-time PCR confirmed induction of EYFP. We successfully replicated the TReATS method in PCLS from Vangl2flox/flox mice, leading to the deletion of loxP-flanked exon 4 of the Vangl2 gene. Cre-treated Vangl2flox/flox PCLS exhibited cytoskeletal abnormalities, a known phenotype caused by VANGL2 dysfunction. We report a novel method that by-passes conventional Cre-Lox breeding, allowing rapid and highly effective gene manipulation in ex vivo tissue models.© 2023. Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributedJournal Articlehttps://www.dx.doi.org/10.1242/dmm.050267https://journals.biologists.com/dmm/article/16/11/dmm050267/334533/A-method-for-TAT-Cre-recombinase-mediated-floxed