Recipient Organization
UNIVERSITY OF ILLINOIS
2001 S. Lincoln Ave.
URBANA,IL 61801
Performing Department
(N/A)
Non Technical Summary
(N/A)
Animal Health Component
40%
Research Effort Categories
Basic
40%
Applied
40%
Developmental
20%
Goals / Objectives
To define the spatial transcriptome of host and virus RNA transcripts within individual cells in the skin of chicken infected with gallid ahphaherpesvirus-2.
Project Methods
Chickens will be infected with wildtype and transmission mutants of Marekâ¿¿s disease virus, and at late times postinoculation, skin biopsies will be collected and frozen. Four tissue samples will be fixed onto cell capture slides that contain barcoded capture oligonucleotides. Once fixed, the tissue sample will be stained with either H&E or IF staining and microscopically imaged. Then, the tissue will be permeabilized to release the mRNA so it can be captured with the 5,000 barcoded capture oligonucleotides that bind to the RNA in the tissue. Finally, the captured mRNA will be converted into cDNA, and sequencing libraries will be prepared. Each barcoded oligonucleotide will capture the transcripts from 1-10 cells. The Visium spatial gene expression platform will link both host and viral transcripts to a specific location on the slide, allowing a visual depiction of the transcriptional landscape.