At the same time, homologous recombination is generally as highthroughput- compatible as LIC. The modified primer design of the yeast-based DREAM mutagenesis method is simple and like the classic SDM covers all aspects of mutagenesis: mutation, insertion and deletion. The DREAM mutagenesis primer pair can easily be designed by the following basic rules: each oligonucleotide should be around 50 bases long, with 20 bases of 59 primer-toprimer overlap for an efficient recombination of the mutated plasmid ends in yeast, carrying the mutation, deletion or insertion in their middle, and 30 bases of 39-sequence for template annealing. The 20 bp overlap on both ends of the linear mutagenesis product results in efficient homology-based gap repair in S. cerevisiae to form the NQ 301 circular, mutated plasmid. To minimize the occurrence of errors, the number of thermal cycles is restricted to 18 as recommended for the conventional SDM kit. Accordingly, all analyzed DREAM clones so far replicated the template plasmid sequence i.e. apart from the introduced mutation the whole BSEP coding sequence was found to be unchanged. The new mutagenesis method permits the rapid realization of patient-derived BSEP mutations for immediate study in cell cultures. Using DREAM, we could show that a BSEP mutation identified in a patient with progressive familial intrahepatic cholestasis type 2 results in a trafficking defect of the mutant protein that prevents BSEP from being correctly incorporated into the plasma membrane. Future mutations can be generated quickly for their study in mammalian cell lines and/or in vitro on the isolated recombinant protein. This is a major advantage since the realization of for example BSEP mutations was previously a workintensive and time-consuming task. Glycosylation has been shown to be irrelevant for the function of other human ABC Impentamine dihydrobromide transporters expressed in yeast in the past. Furthermore, BSEP expressed in Sf9 cells, which also harbors a glycosylation pattern different from the human pattern, was functional. Thus, it is very likely that the glycosylation state and/or pattern of BSEP is not relevant for its function. Sepsis and septic shock involve dysregulated inflammatory responses caused by interaction between the host immune system and microorganisms. Despite recent progress in care, sepsis and septic shock remain associated with high morbidity and mortality, as well as diminished organ function or failure, including the kidneys, lungs, and bone marrow.