Fig. 5.

Strategy for the making of redesigned HEs. a General strategy. I-CreI variant libraries with locally altered specificity are generated. These mutants were assembled into homodimeric and heterodimeric proteins using a combinatorial approach, generating meganucleases with fully redesigned specificity. b The RAG1 series of targets. Two intermediary palindromic targets were derived from the non-palindromic human gene target. These were used to select for the homodimeric I-CreI variants that served as scaffold for the specific heterodimer. The two 3 bp segments used in the library screening are boxed