Systematic analysis of splicing defects in selected primary immunodeficiencies-related genes

Clin Immunol. 2017 Jul:180:33-44. doi: 10.1016/j.clim.2017.03.010. Epub 2017 Mar 27.

Abstract

Both variants affecting splice sites and those in splicing regulatory elements (SREs) can impair pre-mRNA splicing, eventually leading to severe diseases. Despite the availability of many prediction tools, prognosis of splicing affection is not trivial, especially when SREs are involved. Here, we present data on 92 in silico-/55 minigene-analysed variants detected in genes responsible for the primary immunodeficiencies development (namely BTK, CD40LG, IL2RG, SERPING1, STAT3, and WAS). Of 20 splicing-affecting variants, 16 affected splice site while 4 disrupted potential SRE. The presence or absence of splicing defects was confirmed in 30 of 32 blood-derived patients' RNAs. Testing prediction tools performance, splice site disruptions and creations were reliably predicted in contrast to SRE-affecting variants for which just ESRseq, ΔHZEI-scores and EX-SKIP predictions showed promising results. Next, we found an interesting pattern in cryptic splice site predictions. These results might help PID-diagnosticians and geneticists cope with potential splicing-affecting variants.

Keywords: Cryptic splice sites; Primary immunodeficiencies; Splicing prediction; Splicing-affecting variants.

MeSH terms

  • Agammaglobulinaemia Tyrosine Kinase
  • Child
  • Child, Preschool
  • Complement C1 Inactivator Proteins / genetics
  • Complement C1 Inhibitor Protein
  • Exons
  • HeLa Cells
  • Hep G2 Cells
  • Humans
  • Immunologic Deficiency Syndromes / genetics*
  • Infant
  • Interleukin Receptor Common gamma Subunit / genetics
  • Mutation
  • Protein-Tyrosine Kinases / genetics
  • RNA Splicing*
  • RNA, Messenger / genetics
  • Recombinant Fusion Proteins / genetics
  • STAT3 Transcription Factor / genetics
  • U937 Cells
  • Wiskott-Aldrich Syndrome Protein / genetics

Substances

  • CD40LIg fusion protein
  • Complement C1 Inactivator Proteins
  • Complement C1 Inhibitor Protein
  • IL2RG protein, human
  • Interleukin Receptor Common gamma Subunit
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • SERPING1 protein, human
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • WAS protein, human
  • Wiskott-Aldrich Syndrome Protein
  • Protein-Tyrosine Kinases
  • Agammaglobulinaemia Tyrosine Kinase
  • BTK protein, human