Sf3b1^tm1.1Gsva

Using gateway and recombineering recombination technologies, exon 15 was replaced with a copy with an A>G nucleotide substitution changing Lys700 codon AAA to glutamic acid codon GAA. At the same time a loxP site flanked cDNA splice-trap cassette and an FRT site flanked neomycin resistance gene cassette were inserted into intron 11. The cDNA splice-trap cassette comprised the following: an engrailed 2 intron with strong splice acceptor, a partial cDNA for the gene coding for exon 12 onwards (and including introns 15 and 19 to prevent bacterial toxicity), followed by an SV40 pA signal. The neo cassette was removed through subsequent flp-mediated recombination, leaving a conditional-ready K700E mutation allele. The K700E mutation is the most common mutation associated with MDS (age-related clonal hematopoiesis and myelodysplastic syndromes). (J:239625)