Abstract
Retroviral transduction is commonly used to modulate gene expression and is a powerful approach to understand the role of a gene using gain- or loss-of-function strategies. Retroviral vectors can stably integrate non-viral genes into host genomes, providing long-term modulation of gene expression in infected cells and their progeny. Here we describe the generation of retroviral supernatants and the steps to efficiently transduce genes in innate lymphoid cell (ILC) progenitors for subsequent analysis of ILC populations in vivo.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Artis D, Spits H (2015) The biology of innate lymphoid cells. Nature 517:293–301
Serafini N, Vosshenrich CA, Di Santo JP (2015) Transcriptional regulation of innate lymphoid cell fate. Nat Rev Immunol 15:415–428
Mjösberg J, Bernink J, Golebski K et al (2012) The transcription factor GATA3 is essential for the function of human type 2 innate lymphoid cells. Immunity 37:649–659
Seillet C, Huntington ND, Gangatirkar P et al (2014) Differential requirement for Nfil3 during NK cell development. J Immunol 192:2667–2676
Yu X, Wang Y, Deng M et al (2014) The basic leucine zipper transcription factor NFIL3 directs the development of a common innate lymphoid cell precursor. elife 3
van de Pavert SA, Ferreira M, Domingues RG et al (2014) Maternal retinoids control type 3 innate lymphoid cells and set the offspring immunity. Nature 508:123–127
Xu W, Domingues RG, Fonseca-Pereira D et al (2015) NFIL3 orchestrates the emergence of common helper innate lymphoid cell precursors. Cell Rep 10:2043–2054
Possot C, Schmutz S, Chea S et al (2011) Notch signaling is necessary for adult, but not fetal, development of RORγt(+) innate lymphoid cells. Nat Immunol 12:949–958
Klose CSN, Flach M, Möhle L et al (2014) Differentiation of type 1 ILCs from a common progenitor to all helper-like innate lymphoid cell lineages. Cell 157:340–356
Constantinides MG, McDonald BD, Verhoef PA et al (2014) A committed precursor to innate lymphoid cells. Nature 508:397–401
Seillet C, Mielke LA, Amann-Zalcenstein DB et al (2016) Deciphering the innate lymphoid cell transcriptional program. Cell Rep 17:436–447
Seillet C, Rankin LC, Groom JR et al (2014) Nfil3 is required for the development of all innate lymphoid cell subsets. J Ex Med 211:1733–1740
Heinen AP, Wanke F, Moos S et al (2014) Improved method to retain cytosolic reporter protein fluorescence while staining for nuclear proteins. Cytometry A 85:621–627
Acknowledgments
This work was supported by grants (Grants 1122277, 1098832) and fellowships from the National Health and Medical Research Council of Australia (CS and GTB) and the Australian Research Council (GTB). This study was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC Independent Research Institute Infrastructure.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Seillet, C., Belz, G.T. (2019). Assessment of Gene Function of Mouse Innate Lymphoid Cells for In Vivo Analysis Using Retroviral Transduction. In: Moll, J., Carotta, S. (eds) Target Identification and Validation in Drug Discovery. Methods in Molecular Biology, vol 1953. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9145-7_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9145-7_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9144-0
Online ISBN: 978-1-4939-9145-7
eBook Packages: Springer Protocols