DNA Recovery From Tape-Lifting Kits: Methodology and Practice.

In the case of suspicious deaths, the technique of 1:1 taping is often used in Belgium. It consists of affixing a large number of adhesive tapes to the body of the victim. It is conventionally aimed at obtaining microtraces (e.g., fibers, hair) and is usually not used for DNA analysis. However, in some cases, DNA analysis of certain areas of interest identified on the 1:1 taping material can offer a last resort solution. The four-step method that is described in this article involves the selection of areas of interest on the body (Step 1), the selection of the corresponding tapes (Step 2), the decontamination of the tapes (Step 3), the selection of areas of interest on the tapes, for DNA sampling (Step 4). The method is illustrated by its successful application in four murder cases. In each case, DNA profiles of good quality could be identified, including profiles of persons different from the victim.

Homologous Transcription Factors DUX4 and DUX4c Associate with Cytoplasmic Proteins during Muscle Differentiation.

Hundreds of double homeobox (DUX) genes map within 3.3-kb repeated elements dispersed in the human genome and encode DNA-binding proteins. Among these, we identified DUX4, a potent transcription factor that causes facioscapulohumeral muscular dystrophy (FSHD). In the present study, we performed yeast two-hybrid screens and protein co-purifications with HaloTag-DUX fusions or GST-DUX4 pull-down to identify protein partners of DUX4, DUX4c (which is identical to DUX4 except for the end of the carboxyl terminal domain) and DUX1 (which is limited to the double homeodomain). Unexpectedly, we identified and validated (by co-immunoprecipitation, GST pull-down, co-immunofluorescence and in situ Proximal Ligation Assay) the interaction of DUX4, DUX4c and DUX1 with type III intermediate filament protein desmin in the cytoplasm and at the nuclear periphery. Desmin filaments link adjacent sarcomere at the Z-discs, connect them to sarcolemma proteins and interact with mitochondria. These intermediate filament also contact the nuclear lamina and contribute to positioning of the nuclei. Another Z-disc protein, LMCD1 that contains a LIM domain was also validated as a DUX4 partner. The functionality of DUX4 or DUX4c interactions with cytoplasmic proteins is underscored by the cytoplasmic detection of DUX4/DUX4c upon myoblast fusion. In addition, we identified and validated (by co-immunoprecipitation, co-immunofluorescence and in situ Proximal Ligation Assay) as DUX4/4c partners several RNA-binding proteins such as C1QBP, SRSF9, RBM3, FUS/TLS and SFPQ that are involved in mRNA splicing and translation. FUS and SFPQ are nuclear proteins, however their cytoplasmic translocation was reported in neuronal cells where they associated with ribonucleoparticles (RNPs). Several other validated or identified DUX4/DUX4c partners are also contained in mRNP granules, and the co-localizations with cytoplasmic DAPI-positive spots is in keeping with such an association. Large muscle RNPs were recently shown to exit the nucleus via a novel mechanism of nuclear envelope budding. Following DUX4 or DUX4c overexpression in muscle cell cultures, we observed their association with similar nuclear buds. In conclusion, our study demonstrated unexpected interactions of DUX4/4c with cytoplasmic proteins playing major roles during muscle differentiation. Further investigations are on-going to evaluate whether these interactions play roles during muscle regeneration as previously suggested for DUX4c.

Evaluation of novel forensic DNA storage methodologies.

An issue in forensic sciences is the secure storage of extracted DNA. Most of the time, DNA is frozen at -20°C or -80°C. Recently, new room temperature DNA storage technologies have been developed based on anhydrobiosis. Two products use this technology: Qiasafe (Qiagen) and Gentegra (Genvault). In this study we focused on the recent Gentegra product and initiated a comparison versus -20°C and Qiasafe storage. We compared the quantity and quality of DNA stored using anhydrobiosis technology against DNA stored at -20°C, by performing STR profiling after short term storage. Furthermore, we studied the quantity and integrity of DNA after long term storage. Our results prove the high potential of this technology but it seems to be extraction dependent. Phenol/chloroform extracted DNA could be stored using the Gentegra matrix for more than 6 months without any obvious degradation. However, DNA extracted using magnetic beads could not be safely stored over the same period. Adaptations are therefore required to store this kind of samples.