General information
The SALSA MLPA Probemix P479 TCF12 - ERF is a research use only (RUO) assay for the detection of deletions or duplications in the TCF12 and ERF genes, which are associated with craniosynostosis.

Craniosynostosis is a clinically and genetically heterogeneous condition with a prevalence of ~1 in 2200 births. Several genes have been identified as a cause of craniosynostosis including FGFR1, FGFR2, FGFR3, TWIST1, EFNB1, MSX2, ALX4, ZIC1, SMAD6 and, more recently, TCF12 and ERF. Craniosynostosis is inherited in an autosomal dominant manner and is characterised by abnormal skull growth involving premature fusion of the cranial sutures. As a result, the growth velocity of the skull often cannot match that of the developing brain. This produces skull deformity and, in some cases, raises intracranial pressure, which must be treated promptly to avoid permanent neurodevelopmental disability. More information on TCF12-related and ERF-related craniosynostosis can be found in OMIM #615314 and #600775, respectively.

The TCF12 gene spans ~371 kb of genomic DNA and is located on 15q21.3, 55 Mb from the p-telomere. The TCF12 gene encodes a transcriptional regulator that belongs to the basic helix-loop-helix family. Approximately 10% of all craniosynostoses is caused by heterozygous point mutations in the TCF12 gene (Sharma et al. 2013). Large rearrangements involving the complete TCF12 gene have been described. Intragenic deletions and a duplication in the TCF12 gene were reported in craniosynostosis patients (Goos et al. 2016).

The ERF gene spans ~8 kb of genomic DNA and is located on 19q13.2, 47 Mb from the p-telomere. The ERF gene encodes a transcriptional repressor that belongs to the ETS transcription factors family. No mutations or rearrangements involving the ERF gene have been reported so far. However, it has been shown that reduced dosage of ERF causes complex craniosynostosis in humans and mice (Twigg et al. 2013).

This SALSA MLPA probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.

Probemix content
The SALSA MLPA Probemix P479-A1 TCF12 - ERF contains 40 MLPA probes with amplification products between 130 and 452 nucleotides (nt). This includes 25 probes for the TCF12 gene (one probe for each exon and two probes for exons 4, 5, 6 and 8) and five probes for the ERF gene (one probe for each exon and one probe for intron 1). In addition, ten reference probes are included that detect autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes are available online (www.mrcholland.com).

This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at www.mrcholland.com.