Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice

• Type of Material: Article
• Authors: Lauren E. Hudson, Emory University; Taryn P. Stewart, Emory University; Milo Fasken, Emory University; Anita Corbett, Emory University; Tracey Lamb, Emory University
• Language: English
• Date: 2016-02-01
• Publisher: Journal of Visualized Experiments (JoVE)
• Publication Version: Final Published Version
• Copyright Statement: © 2016 Journal of Visualized Experiments.
• Final Published Version (URL): http://dx.doi.org/10.3791/53453
• Title of Journal or Parent Work: Journal of Visualized Experiments
• ISSN: 1940-087X
• Volume: 2016
• Issue: 108
• Start Page: e53453
• End Page: e53453
• Grant/Funding Information: The authors acknowledge funding through the Children's Center for Immunology and Vaccines and an NIH New Innovator Award (1DP2AI112242-01) awarded to Tracey J. Lamb.
• Keywords: heterologous protein; auxotroph; Saccharomyces boulardii; gavage; probiotic; GFP; URA3; UV mutagenesis; Multidisciplinary Sciences; Peyer's patch; Science & Technology; Issue 108; Immunology
• Research Categories: Health Sciences, Immunology; Chemistry, Biochemistry

Abstract

Development of recombinant oral therapy would allow for more direct targeting of the mucosal immune system and improve the ability to combat gastrointestinal disorders. Adapting probiotic yeast in particular for this approach carries several advantages. These strains have not only the potential to synthesize a wide variety of complex heterologous proteins but are also capable of surviving and protecting those proteins during transit through the intestine. Critically, however, this approach requires expertise in many diverse laboratory techniques not typically used in tandem. Furthermore, although individual protocols for yeast transformation are well characterized for commonly used laboratory strains, emphasis is placed here on alternative approaches and the importance of optimizing transformation for less well characterized probiotic strains. Detailing these methods will help facilitate discussion as to the best approaches for testing probiotic yeast as oral drug delivery vehicles and indeed serve to advance the development of this novel strategy for gastrointestinal therapy.