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ARS Home » Plains Area » Clay Center, Nebraska » U.S. Meat Animal Research Center » Genetics and Animal Breeding » Research » Publications at this Location » Publication #364273

Research Project: Identifying Genomic Solutions to Improve Efficiency of Swine Production

Location: Genetics and Animal Breeding

Title: Deciphering the dynamics of porcine circovirus associated diseases using gene silencing and editing

Author
item WALKER, LIANNA - UNIVERSITY OF NEBRASKA
item ENGLE, TAYLOR - UNIVERSITY OF NEBRASKA
item VU, HIEP - UNIVERSITY OF NEBRASKA
item TOSKY, EMILY - UNIVERSITY OF NEBRASKA
item Nonneman, Danny - Dan
item Smith, Timothy - Tim
item BORZA, TUDOR - DALHOUSIE UNIVERSITY
item BURKEY, THOMAS - UNIVERSITY OF NEBRASKA
item PLASTOW, GRAHAM - UNIVERSITY OF ALBERTA
item KACHMAN, STEPHEN - UNIVERSITY OF NEBRASKA
item 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., Engle, T., Vu, H., Tosky, E., Nonneman, D.J., Smith, T.P.L., Borza, T., Burkey, T., Plastow, G.S., Kachman, S.D., Ciobanu, D.C. 2019. Deciphering the dynamics of porcine circovirus associated diseases using gene silencing and editing [abstract]. In: Plant and Animal Genome Conference Proceedings XXVII, 12-16 Jan. 2019, San Diego, CA. pg. 216, W1045. 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 the SYNGR2 gene. In vitro silencing of SYNGR2 in PK15 cells via siRNA transfection resulted in a onelog reduction in PCV2b 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 SYNGR2 generated a PK15 edited clone homozygous for a 106bp deletion within the second exon that exhibited a two-log reduction in PCV2b titer following infection compared to wild-type PK15 cells (P<0.05). Given these findings and that SYNGR2 p.Arg63Cys is the only missense mutation within this gene, SYNGR2 p.Arg63Cys is a plausible QTN for PCV2b susceptibility.