. / The investigators 1) created induced pluripotent stem cells (iPSCs) from patients with PiZZ-associated A1AT deficiency; 2) designed zinc finger nucleases to specifically nick the DNA flanking the mutation in the A1AT gene; 3) replaced the region of genomic DNA containing the mutation with a fragment of DNA containing not only the corrected gene sequence, but also a transposon, which allows for selective growth of corrected iPSCs; 4) removed the transposon without leaving a trace; 5) assessed for corrected clones that had not undergone additional mutations during the procedures using deep sequencing, genome hybridization, and other approaches; and 6) differentiated the iPSCs into functional hepatocytes that were transplanted into immunodeficient mice, where they functioned to produce human albumin and the corrected A1AT protein.

The investigators 1) created induced pluripotent stem cells (iPSCs) from patients with PiZZ-associated A1AT deficiency; 2) designed zinc finger nucleases to specifically nick the DNA flanking the mutation in the A1AT gene; 3) replaced the region of genomic DNA containing the mutation with a fragment of DNA containing not only the corrected gene sequence, but also a transposon, which allows for selective growth of corrected iPSCs; 4) removed the transposon without leaving a trace; 5) assessed for corrected clones that had not undergone additional mutations during the procedures using deep sequencing, genome hybridization, and other approaches; and 6) differentiated the iPSCs into functional hepatocytes that were transplanted into immunodeficient mice, where they functioned to produce human albumin and the corrected A1AT protein.

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