September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Exome Analysis Detected a Novel 7.4 kb Deletion in the PRPH2 Gene as the Underlying Cause of Recessive Retinal Degeneration in a Consanguineous Family.
Author Affiliations & Notes
  • Kameron Kishaba
    Shiley Eye Institute, University of California, San Diego, La Jolla, California, United States
  • Pooja Biswas
    Shiley Eye Institute, University of California, San Diego, La Jolla, California, United States
  • John Suk
    Shiley Eye Institute, University of California, San Diego, La Jolla, California, United States
  • Inayat Ullah
    National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
  • Muhammad Asif Naeem
    National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
  • Sheikh Riazuddin
    National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
    Allama Iqbal Medical College, University of Health Sciences, Lahore, Pakistan
  • Javed Akram
    Allama Iqbal Medical College, University of Health Sciences, Lahore, Pakistan
    National Centre for Genetic Diseases, Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad, Pakistan
  • J. Fielding Hejtmancik
    OMGS/OGVFB, National Eye Institute, NIH, Bethesda, Maryland, United States
  • S Amer Riazuddin
    The Wilmer Eye Institute, John's Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Radha Ayyagari
    Shiley Eye Institute, University of California, San Diego, La Jolla, California, United States
  • Footnotes
    Commercial Relationships   Kameron Kishaba, None; Pooja Biswas, None; John Suk, None; Inayat Ullah, None; Muhammad Naeem, None; Sheikh Riazuddin, None; Javed Akram, None; J. Fielding Hejtmancik, None; S Amer Riazuddin, None; Radha Ayyagari, None
  • Footnotes
    Support  Foundation Fighting Blindness, Research to Prevent Blindness, P30EY022589, NIH-EY13198, NIH-EY21237
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 3137. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Kameron Kishaba, Pooja Biswas, John Suk, Inayat Ullah, Muhammad Asif Naeem, Sheikh Riazuddin, Javed Akram, J. Fielding Hejtmancik, S Amer Riazuddin, Radha Ayyagari; Exome Analysis Detected a Novel 7.4 kb Deletion in the PRPH2 Gene as the Underlying Cause of Recessive Retinal Degeneration in a Consanguineous Family.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3137.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : To identify the causative mutation in an inbred large Pakistani pedigree with recessive retinal degeneration (RD).

Methods : Ophthalmic evaluation included measurement of visual acuity, color vision, fundus examination, and full-field electroretinography (ERG). Blood samples were collected from six affected, and nine unaffected individuals, and genomic DNA was isolated using standard protocols. Next generation whole exome libraries were prepared using Agilent V5+UTRs capture kit, which were sequenced on Illumina HiSeq2000. The sequence variants were filtered by exomeSuite using an autosomal recessive inheritance model. Homozygosity mapping was performed using exomeSuite to identify homozygous regions shared by affected members, which were screened for the presence of known retinal disease genes. Examination of candidate gene sequence reads pile-up was carried out for large deletions or rearrangements. Mapping the boundaries of the detected deletion was performed by PCR and sequencing, while segregation analysis was performed by PCR using three primers that amplify the wild type and mutant alleles.

Results : An eight-generation Pakistani family consisting of ten affected individuals and seven consanguineous marriages was recruited. Affected individuals were diagnosed with retinal degeneration. Whole exome analysis detected 23,695 homozygous variants shared by six affected members of the pedigree. Examination of these variants did not identify potential causative variants. Homozygosity mapping revealed three homozygous regions on chromosomes 5, 6, and 9 harboring three known RD genes, GUCA1B, PRPH2 and TLR4. Among these, PRPH2 gene on chromosome 6 has been reported to be associated with dominant and recessive RD. In this pedigree, PCR amplification and examination of exome sequence reads pile-up of the PRPH2 gene revealed a homozygous deletion encompassing the exon 2 of this gene. Amplification and sequencing the IVS1, IVS2 and exon2 of PRPH2 detected a deletion of 7408 base pairs (c.582-2420_c.828+4741del) spanning from IVS1 to IVS2. The deletion segregated with the RD phenotype in this pedigree and identified six unaffected heterozygous carriers.

Conclusions : Analysis of exome sequence variants and homozygosity mapping revealed a novel 7408bp homozygous deletion in the PRPH2 segregating with recessive RD.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×