Biotechnology tools and challenges A NEW STRATEGY TO IDENTIFY THE DISEASE CAUSING MUTATION FOR NEURONAL CEROID LIPOFUSCINOSIS IN SOUTH HAMSPHIRE SHEEP

The New Zealand South Hampshire sheep have been well characterised as an animal model for variant late-infantile neuronal ceroid lipofuscinosis (vLINCL) in humans. The disease causing gene has been identified as CLN6, but so far no mutation has been identified. A sheep BAC containing CLN6 and flanking region (~120kb) was sequenced at 13.49-fold sequence coverage using the 454 sequencing method from Roche® to complement the existing but incomplete public domain ovine sequence for the region of interest. The 454 sequence was assembled to bovine genomic sequence on chromosome 10 (BTA10). For mutation screening 15 long-distance PCR products from affected and normal South Hampshire sheep covering CLN6 as well as substantial 3’ and 5’ flanking sequence are currently optimized to be sequenced. Regulatory elements and/or mutations identified in CLN6 non-coding regions are likely to indicate positions for disease causing mutations not only in the South Hampshire sheep but also in human uncharacterised vLINCL patients.