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Scoop.it!
The development of programmable nucleases has enabled the application of new genome engineering strategies for cellular immunotherapy. 
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Scoop.it!
BigField GEG Tech's insight:
The authors present a protocol for using ΦC31 and Bxb1 ntegrases simultaneously to facilitate cassette exchange at a predefined location. This approach permits greater control and accuracy over integration. They also present a general method for using polymerase chain reaction assays to verify that the desired cassette exchange occurred successfully.
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Here the authors show a proof-of-concept for reprogramming MHC-specificity by performing CRISPR-Cas9-assisted cassette exchange. Using murine antigen presenting cell lines (RAW264.7 macrophages), they demonstrate that the generation of Cas9-induced double-stranded breaks flanking the native MHC-I H2-Kd locus led to exchange of an orthogonal H2-Kb allele. These findings highlight a potential new approach for the correcting of MHC mismatches in cellular transplantation.