Journal article

Transient, flexible gene editing in zebrafish neutrophils and macrophages for determination of cell-autonomous functions

Abdulsalam I Isiaku, Zuobing Zhang, Vahid Pazhakh, Harriet R Manley, Ella R Thompson, Lucy C Fox, Satwica Yerneni, Piers Blombery, Graham J Lieschke

DISEASE MODELS & MECHANISMS | COMPANY BIOLOGISTS LTD | Published : 2021

Abstract

Zebrafish are an important model for studying phagocyte function, but rigorous experimental systems to distinguish whether phagocyte-dependent effects are neutrophil or macrophage specific have been lacking. We have developed and validated transgenic lines that enable superior demonstration of cell-autonomous neutrophil and macrophage genetic requirements. We coupled well-characterized neutrophil- and macrophage-specific Gal4 driver lines with UAS:Cas9 transgenes for selective expression of Cas9 in either neutrophils or macrophages. Efficient gene editing, confirmed by both Sanger and next-generation sequencing, occurred in both lineages following microinjection of efficacious synthetic guid..

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Grants

Awarded by China Scholarship Council


Awarded by Maddie Riewoldt's Vision


Awarded by National Health and Medical Research Council


Awarded by Australian Research Council


Funding Acknowledgements

This work was supported by Monash University (Graduate Scholarship and International Tuition Scholarship to A.I.I.; Graduate Scholarship and International Postgraduate Research Fellowship to V.P.; Research Training Program Stipend and Postgraduate Publication Award to H.R.M.) , China Scholarship Council (201608140011 to Z.Z.) , Maddie Riewoldt's Vision (MRV2018DM to L.C.F.; ARMI-MRV-2018G to G.J.L.) , National Health and Medical Research Council (1044754, 1086020, 1159278 to G.J.L.) and the Australian Research Council (DP170102235 to G.J.L.) . The Sir Peter MacCallum Cancer Centre Molecular Hematology Laboratory is supported by funding from Wilson Centre for Lymphoma Genomics through the Snowdome Foundation. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government.