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| Description: Humanized sickle cell disease mouse model (BS/BS) 9 Transplant corrected ‘ , 0 Harvest tail hematopoietic cells back ' 'x ._ fibroblasts. into irradiated mice, ' w . Grow fibroblasts thus curing them of in culture. sickle cell disease. Can follow HP cells with GFP [35/135 _ fibroblasts B‘IB‘ HP cells 9 De—differentiate fibroblasts into induced pluripotent . stem (iPS) cells; infect with o Infcect .w'th Moloney three Moloney retroviruses So;%2:25l:::gr::sc;nmgote expressing Oct4, 50x2, and differentiation 0f iPS BS/Bs filnfiigifasesexacgsg if c-M c cells into B‘IB‘ iPS cells p _g y hematopoietic iPS cells 35/35 gene (c-Myc gene '5 flanked progenitor (HP) cells. iPS ““5 by IOXP Sites). //\ y \y "I, / /’ \\\ Infect with adenovirus express- / . . . ‘\\ ing Cre recombinase to /// e Eglggéirggsg‘g‘elzome ‘\\\ remove c-Myc from iPS cells. // editing (below). ‘\\ NZ 9’70, ‘3’” 13"? 5’73 7 .ay 7, 3.. E [35 mouse genomic DNA Mouse DNA Human DNA ay2 BhO Bh1%[3h2 Bh3 ay [35 IB‘ mutation ’, “ “ ‘“ ’ “v ‘P ‘9”, | [Sites targeted by guide RNAs CRISPR—CasS induces ds breaks. 1 ay2 BhO fih1 [3h2 Bh3 ay (.5 Repair via homologous recombination with supplied 1 inner DNA fragments containing wild~type BAallele. ayZ [3h0 [3h1 BhZ Bh3 ay [3‘
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