Engineering Disease Resistance through Generation of an iPSC-derived P. Maniculatus Chimera

Abstract

The goal of this project is to develop methods to manipulate the genome of Peromyscus maniculatus for the future eradication of diseases such as Hantavirus and Lyme disease that this species can transmit to humans. The initial thrust of this project is to generate induced pluripotent stem cells (iPSCs) from P. maniculatus mouse embryonic fibroblasts (MEFs) using an engineered Sendai Virus vector system. The Sendai Virus vector is a non-integrating viral vector that delivers four transcription factors necessary to reprogram normal somatic cells to a pluripotent, stem cell-like state. Subsequent, a plasmid will be used to insert DNA into the genome of the iPSC that codes for a fluorescent protein. In the future, the fluorescence will be used to confirm that Mus musculus cells are able to combine with the P. maniculatus cells in a chimeric blastocyst. Because many notable diseases are transmissible to humans from P. maniculatus, the genetic manipulation and creation of a disease resistant P. maniculatus using iPSCs could help to eradicate the spread of these diseases.