Improved full-length killer cell immunoglobulin-like receptor transcript discovery in Mauritian cynomolgus macaques.

TitleImproved full-length killer cell immunoglobulin-like receptor transcript discovery in Mauritian cynomolgus macaques.
Publication TypeJournal Article
Year of Publication2017
AuthorsPrall, TM, Graham, ME, Karl, JA, Wiseman, RW, Ericsen, AJ, Raveendran, M, Harris, RA, Muzny, DM, Gibbs, RA, Rogers, J, O'Connor, DH
Date Published2017 05
KeywordsAnimals, Exome, Gene Frequency, High-Throughput Nucleotide Sequencing, Macaca fascicularis, Receptors, KIR, Sequence Analysis, DNA

Killer cell immunoglobulin-like receptors (KIRs) modulate disease progression of pathogens including HIV, malaria, and hepatitis C. Cynomolgus and rhesus macaques are widely used as nonhuman primate models to study human pathogens, and so, considerable effort has been put into characterizing their KIR genetics. However, previous studies have relied on cDNA cloning and Sanger sequencing that lack the throughput of current sequencing platforms. In this study, we present a high throughput, full-length allele discovery method utilizing Pacific Biosciences circular consensus sequencing (CCS). We also describe a new approach to Macaque Exome Sequencing (MES) and the development of the Rhexome1.0, an adapted target capture reagent that includes macaque-specific capture probe sets. By using sequence reads generated by whole genome sequencing (WGS) and MES to inform primer design, we were able to increase the sensitivity of KIR allele discovery. We demonstrate this increased sensitivity by defining nine novel alleles within a cohort of Mauritian cynomolgus macaques (MCM), a geographically isolated population with restricted KIR genetics that was thought to be completely characterized. Finally, we describe an approach to genotyping KIRs directly from sequence reads generated using WGS/MES reads. The findings presented here expand our understanding of KIR genetics in MCM by associating new genes with all eight KIR haplotypes and demonstrating the existence of at least one KIR3DS gene associated with every haplotype.

Alternate JournalImmunogenetics
PubMed ID28343239
PubMed Central IDPMC5856007
Grant ListC06 RR020141 / RR / NCRR NIH HHS / United States
C06 RR015459 / RR / NCRR NIH HHS / United States
HHSN272201100013I / AI / NIAID NIH HHS / United States
T32 AI078985 / AI / NIAID NIH HHS / United States
R01 AI116382 / AI / NIAID NIH HHS / United States
HHSN272201600007C / AI / NIAID NIH HHS / United States
HHSN272201100013C / AI / NIAID NIH HHS / United States

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