This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To introduce a selectable marker such as neomycin resistance or EGFP by homologous recombination into a tissue specific gene. We introduced EGFP and puromycin into hSox1 located on chromosome 13q34 via homologous recombination. Eight clones were obtained. When these hSox1 knock in cells underwent neural differentiation low levels of EGFP mRNA expression was observed via RT-PCR. We plan to optimize the neural differentiation protocol and purify hSox1 cells to determine their developmental potential and analyze their gene expression. This research uses WNPRC Stem Cell Resources and federally approved human ES cell lines.
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