Technical Details:
Multiple myeloma (MM) is an incurable hematologic malignancy characterized by recurrent chromosomal translocations. Patients with t(4;14)(p16;q32) are the worst prognostic subgroup in MM, although the basis for this poor prognosis is unknown. The t(4;14) involves 2 potential target genes: fibroblast growth factor receptor 3 (FGFR3) and multiple myeloma SET domain (MMSET). JHU Scientist demonstrate a role for MMSET in t(4;14) and additionally develop a MMSET Knockout/Knockdown cell line. JHU scientists have developed a Multiple Myeloma knock out/knock down cell line where the MMSET gene was knocked out and as a control the non-translocated allele was also knocked out. The MMSET gene was disrupted in KMS-11 cells by homologous recombination. After Cre-loxP recombination a single loxP site is left in place of the deleted exon 7. Splicing from exon 6 to exon 8 results in a reading frame shift and premature termination of translation after exon 6 (MMSET T-KO). Immunoblotting demonstrates extensive depletion of full length MMSET protein in TKO cells but not in non-T-KO cells.
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MMSET Knockout/Knockdown cell line
