title: Fltp(T2AiCre): a new knock-in mouse line for conditional gene targeting in distinct mono- and multiciliated tissues.
abstract: We recently identified Flattop (Fltp; 1700009p17Rik) in a screen for potential Foxa2 target and novel mouse organizer genes. Besides its expression in the embryonic node, we found that Fltp is active in other monociliated tissues such as the sensory organs of the inner ear, duct and islets of the pancreas as well as in testis. Additionally, Fltp mRNA is expressed in multiciliated epithelial cells of the lung and of the choroid plexi in the brain. To genetically lineage trace these cells during development and injury as well as to conditionally inactivate genes in these tissues, we generated a Cre recombinase knock-in mouse line using the Fltp gene locus. By homologous recombination we have fused the Fltp open-reading frame to a tandem affinity purification (TAP) tag followed by an intervening viral T2A sequence for co-translational cleavage and an improved Cre recombinase (iCre). This strategy allows both the analysis of the tagged Fltp-TAP-T2A protein and the usage of the iCre recombinase for conditional targeting approaches. Using the ROSA26 reporter mouse line we show that Fltp(T2AiCre) is first active in the monociliated cells of the node, notochord, floorplate and prechordal plate, consistent with the Fltp-TAP-T2A protein production in the node progenitor cells. Furthermore iCre recombinase activity is detected in multiciliated tissues such as choroid plexi of the brain and epithelial cells of the lung with the onset at E10.5 and E13.5, respectively. In the pancreas, β-galactosidase activity is seen in the monociliated cells of the pancreatic duct and islet of Langerhans. Intercrossing Fltp(T2AiCre) mice with the CAG-CAT-EGFP reporter mouse line further confirms iCre activity in multiciliated cells of the lung and brain on a cellular level. Thus, the Fltp(T2AiCre) line is a powerful tool to conditionally inactivate genes in distinct mono- and multiciliated tissues and to analyze the tagged Fltp protein in vivo.
authors.affiliation: Institute of Diabetes and Regeneration Research, Germany.
authors.forename: Anika
authors.initials: A
authors.lastname: Lange
authors.forename: Moritz
authors.initials: M
authors.lastname: Gegg
authors.forename: Ingo
authors.initials: I
authors.lastname: Burtscher
authors.forename: Doris
authors.initials: D
authors.lastname: Bengel
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.forename: Heiko
authors.initials: H
authors.lastname: Lickert
mesh_terms: D000818:Animals; D002460:Cell Line; D002923:Cilia / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism; D055879:Gene Knock-In Techniques; D018390:Gene Targeting; D005838:Genotype; D007150:Immunohistochemistry; D019426:Integrases / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D010179:Pancreas / Q000378:metabolism
chemical_list: C045073:Cre recombinase; D019426:Integrases