Location: Genetics and Animal Breeding
Title: Host genetics influences replication of porcine circovirus 2bAuthor
WALKER, LIANNA - University Of Nebraska | |
ENGLE, TAYLOR - University Of Nebraska | |
VU, HEIP - University Of Nebraska | |
TOSKY, EMILY - University Of Nebraska | |
Nonneman, Danny - Dan | |
Smith, Timothy - Tim | |
BORZA, TUDOR - Dalhousie University | |
BURKEY, THOMAS - University Of Nebraska | |
PLASTOW, GRAHAM - University Of Alberta | |
KACHMAN, STEPHEN - University Of Nebraska | |
CIOBANU, DANIEL - University Of Nebraska |
Submitted to: Plant and Animal Genome Conference Proceedings
Publication Type: Abstract Only Publication Acceptance Date: 12/1/2018 Publication Date: 1/16/2019 Citation: Walker, L.R., Engle, T.B., Vu, H., Tosky, E.R., Nonneman, D.J., Smith, T.P.L., Borza, T., Burkey, T.E., Plastow, G.S., Kachman, S.D., Ciobanu, D.C. 2019. Host genetics influences replication of porcine circovirus 2b [abstract]. In: Plant and Animal Genome Conference XXVII Proceedings, 12-16 January 2019, San Diego, CA. pg. 233. W1136. Available: https://www.intlpag.org/2019/images/pdf/2019/PAGXXVII-abstracts-workshops.pdf Interpretive Summary: Technical Abstract: Porcine Circovirus 2 (PCV2) is the smallest known DNA virus capable of infecting mammals and the primary agent required for the development of a set of symptoms collectively known as Porcine Circovirus Associated Diseases. However, infection with PCV2 does not guarantee clinical disease with variation in severity observed between breeds and individuals. Experimental infection of ~1,000 pigs with PCV2b and genome-wide association analyses revealed two QTL for PCV2b viral load, located on SSC7 and SSC12. Dissection of the SSC12 QTL using ab initio gene prediction, RNAseq, and genomic sequencing identified 66 novel polymorphisms within and upstream of five positional candidate genes. Single marker association analysis of a subset of pigs with extreme high and low viral loads, identified two novel polymorphisms in high LD accounting for 21-23% of the phenotypic variation. One polymorphism is a missense mutation (p.Arg63Cys) within the second exon and critical domain of a novel gene, coding syg93. In vitro silencing of syg93 in PK15 cells via siRNA transfection resulted in reduced PCV2b viral titer (P<0.05) compared to scramble siRNA and non-transfected control cells, indicating a direct role of this gene in PCV2b infection. Additionally, gene editing using CRISPR-Cas9 ribonucleoprotein complexes targeting the second exon of syg93 generated a PK15 edited clone homozygous for a 106bp deletion that exhibited significantly lower PCV2b viral titer following infection compared to wild-type PK15 cells (P<0.05). Given these findings and that syg93 p.Arg63Cys is the only missense mutation within this gene, syg93 p.Arg63Cys is a plausible QTN for PCV2b susceptibility. |