Alu element in the RNA binding motif protein, X-linked 2 (RBMX2) gene found to be linked to bipolar disorder

Objective We have used long-read single molecule, real-time (SMRT) sequencing to fully characterize a similar to 12Mb genomic region on chromosome Xq24-q27, significantly linked to bipolar disorder (BD) in an extended family from a genetic sub-isolate. This family segregates BD in at least four generations with 24 affected individuals. Methods We selected 16 family members for targeted sequencing. The selected individuals either carried the disease haplotype, were non-carriers of the disease haplotype, or served as married-in controls. We designed hybrid capture probes enriching for 5-9Kb fragments spanning the entire 12Mb region that were then sequenced to screen for candidate structural variants (SVs) that could explain the increased risk for BD in this extended family. Results Altogether, 201 variants were detected in the critically linked region. Although most of these represented common variants, three variants emerged that showed near-perfect segregation among all BD type I affected individuals. Two of the SVs were identified in or near genes belonging to the RNA Binding Motif Protein, X-Linked (RBMX) gene family-a 330bp Alu (subfamily AluYa5) deletion in intron 3 of the RBMX2 gene and an intergenic 27bp tandem repeat deletion between the RBMX and G protein-coupled receptor 101 (GPR101) genes. The third SV was a 50bp tandem repeat insertion in intron 1 of the Coagulation Factor IX (F9) gene. Conclusions Among the three genetically linked SVs, additional evidence supported the Alu element deletion in RBMX2 as the leading candidate for contributing directly to the disease development of BD type I in this extended family.Peer reviewe

Phylogenetic Relationships of the Cuscuses and Brushtail Possums (Marsupialia: Phalangeridae) Using the Nuclear Gene BRCA1

The family Phalangeridae comprises approximately two dozen extinct and extant species that include the brushtail possums (Trichosurus), scaly-tailed possum (Wyulda) and cuscuses (Phalanger, Strigocuscus, Spilocuscus and Ailurops). Morphological studies have suggested that Ailurops ursinus is the sister taxon to all other phalangerids. Another species of interest is Strigocuscus celebensis, whose morphologically based taxonomic affinity has habitually been with trichosurins. Mitochondrial 12S rRNA results, however, found moderate support for an Ailurops and Strigocuscus celebensis clade and placed A. ursinus and S. celebensis as sister to Phalanger and Spilocuscus. This study uses nuclear sequence data from the breast cancer and ovarian cancer susceptibility gene 1 (BRCA1) to test previous mitochondrial DNA results and uses relaxed molecular clock methods to estimate divergence dates. The results support Ailurops as sister taxon to S. celebensis and this clade as sister to Phalangerini. Relaxed molecular-dating methods suggest a date of 23-29 million years for the split between Trichosurini and the remaining phalangerids and 19-24 million years for the split between Ailurops + Strigocuscus celebensis and Phalangerini. Several vicariant/dispersal events are necessary to explain the geographic distribution of the Phalangeridae and our estimated molecular divergence dates are congruent with previously proposed south-east Asian geological events

Structural variation and its potential impact on genome instability: Novel discoveries in the EGFR landscape by long-read sequencing.

Structural variation (SV) is typically defined as variation within the human genome that exceeds 50 base pairs (bp). SV may be copy number neutral or it may involve duplications, deletions, and complex rearrangements. Recent studies have shown SV to be associated with many human diseases. However, studies of SV have been challenging due to technological constraints. With the advent of third generation (long-read) sequencing technology, exploration of longer stretches of DNA not easily examined previously has been made possible. In the present study, we utilized third generation (long-read) sequencing techniques to examine SV in the EGFR landscape of four haplotypes derived from two human samples. We analyzed the EGFR gene and its landscape (+/- 500,000 base pairs) using this approach and were able to identify a region of non-coding DNA with over 90% similarity to the most common activating EGFR mutation in non-small cell lung cancer. Based on previously published Alu-element genome instability algorithms, we propose a molecular mechanism to explain how this non-coding region of DNA may be interacting with and impacting the stability of the EGFR gene and potentially generating this cancer-driver gene. By these techniques, we were also able to identify previously hidden structural variation in the four haplotypes and in the human reference genome (hg38). We applied previously published algorithms to compare the relative stabilities of these five different EGFR gene landscape haplotypes to estimate their relative potentials to generate the EGFR exon 19, 15 bp canonical deletion. To our knowledge, the present study is the first to use the differences in genomic architecture between targeted cancer-linked phased haplotypes to estimate their relative potentials to form a common cancer-linked driver mutation