Studies

One of the main characteristics of BLM syndrome patients and BLM deficient cell lines is the enhanced occurence of sister chromatid exhange, detectable in metaphase chromosome spreads. Successful BLM depletion in our homozygous BLM-AID cell line was accompanied by a significant increase of sister chromatid exchange (SCE).

To study how mitotic functions of BLM helicase contribute to genome stability, we genereated chromosomally stable HCT116 cell lines, in which BLM protein can be rapidly degraded in a cell-cycle specific manner. Using the CRISPR/Cas9 technology, a homology donor encoding an auxin-inducible degron (AID), GFP and a selection marker was inserted at the last coding exon of the endogenous BLM gene. To generate the homology donor, a modular cloning system was set up using the Golden Gate assembly strategy. ...