TReATS: a novel method for TAT-Cre recombinase mediated floxed Allele modification in ex vivo tissue slices
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Published version
Author(s)
Type
Journal Article
Abstract
Precision-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.
Date Issued
2023-11
Online Publication Date
2023-11-01T10:45:17Z
Date Acceptance
2023-10-06
ISSN
1754-8403
Publisher
The Company of Biologists
Journal / Book Title
Disease Models and Mechanisms
Volume
16
Issue
11
Copyright Statement
© 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 attributed
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 attributed
License URI
Identifier
https://journals.biologists.com/dmm/article/16/11/dmm050267/334533/A-method-for-TAT-Cre-recombinase-mediated-floxed
Publication Status
Published
Article Number
dmm050267
Date Publish Online
2023-11-03