biostudies-literatureTajer BJNICHD NIH HHSNIGMS NIH HHS21-34https://www.ebi.ac.uk/biostudies/studies/S-EPMC12433595biostudies-literatureUnknown240The axolotl salamander model has broad utility for regeneration studies, but this model is limited by a lack of efficient cell-culture-based tools. The Axolotl Limb-1 (AL-1) fibroblast line, the only available immortalized axolotl cell line, was first published over 20 years ago, but many established molecular biology techniques, such as lipofectamine transfection, CRISPR-Cas9 mutagenesis, and antibiotic selection, work poorly or remain untested in AL-1 cells. Innovating technologies to manipulate AL-1 cells in culture and study their behavior following transplantation into the axolotl will complement in-vivo studies, decrease the number of animals used, and enable the faster, more streamlined investigation of regenerative biology questions. Here, we establish transfection, mutagenesis, anMethods (San Diego, Calif.)Optimized toolkit for the manipulation of immortalized axolotl fibroblasts.PMC12433595R25 GM109436R01 HD095494Fei JFBohm SMin SRoy SSouchet NRWhited JLDecaux AKalu GKarabacak ASousounis KKim RTNelson JABlair SJGilbert PLopez NJCourtemanche KFroitzheim THarake NSavage AMJay SWynn EKidd MDTanaka EMTajer BJPayzin-Dogru DHan JLuong AGSinger HDfalseOptimized toolkit for the manipulation of immortalized axolotl fibroblasts.The axolotl salamander model has broad utility for regeneration studies, but this model is limited by a lack of efficient cell-culture-based tools. The Axolotl Limb-1 (AL-1) fibroblast line, the only available immortalized axolotl cell line, was first published over 20 years ago, but many established molecular biology techniques, such as lipofectamine transfection, CRISPR-Cas9 mutagenesis, and antibiotic selection, work poorly or remain untested in AL-1 cells. Innovating technologies to manipulate AL-1 cells in culture and study their behavior following transplantation into the axolotl will complement in-vivo studies, decrease the number of animals used, and enable the faster, more streamlined investigation of regenerative biology questions. Here, we establish transfection, mutagenesis, an2025-01-01T00:00:00Z2025 Aug2026-06-04T01:34:15.767Z2026-05-04T03:12:22.598ZS-EPMC124335954018738710.1016/j.ymeth.2025.03.019