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The methods and compositions described herein serve to increase the specificity of these novel tools to ensure that the desired target sites will be the primary place of cleavage. The engineered cleavage half domains described can also be used in gene modification protocols requiring simultaneous cleavage at multiple targets either to delete the intervening region or to alter two specific loci at once. Subsequently adding CFTRinh-172, a commercially available small molecule inhibitor of CFTR chloride channel function, lowers the overall conductance indicating chloride ion transport is specific to the CFTR ion channel. ZFNs targeted to sites in the CFTR gene were designed and incorporated into plasmids vectors essentially as described in Urnov et al. At animal sacrifice, Gsalesstaging.Wpengine.Com epithelial cells are isolated and assayed for the amount of CFTR produced and is found in the normal range. Dissociated lung cells were washed, then ran through a column of magnetic beads conjugated to mouse CD45 antibodies (Miltenyi). Then welcome to YesCams

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