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Synthesis of programmable integrases.

ABSTRACT: Accurate modification of the 3 billion-base-pair human genome requires tools with exceptional sequence specificity. Here, we describe a general strategy for the design of enzymes that target a single site within the genome. We generated chimeric zinc finger recombinases with cooperative DNA-binding and catalytic specificities that integrate transgenes with >98% accuracy into the human genome. These modular recombinases can be reprogrammed: New combinations of zinc finger domains and serine recombinase catalytic domains generate novel enzymes with distinct substrate sequence specificities. Because of their accuracy and versatility, the recombinases/integrases reported in this work are suitable for a wide variety of applications in biological research, medicine, and biotechnology where accurate delivery of DNA is desired.

SUBMITTER: Gordley RM

PROVIDER: S-EPMC2654808 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature

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Synthesis of programmable integrases.

Gordley Russell M RM Gersbach Charles A CA Barbas Carlos F CF

Proceedings of the National Academy of Sciences of the United States of America 20090312 13

Accurate modification of the 3 billion-base-pair human genome requires tools with exceptional sequence specificity. Here, we describe a general strategy for the design of enzymes that target a single site within the genome. We generated chimeric zinc finger recombinases with cooperative DNA-binding and catalytic specificities that integrate transgenes with >98% accuracy into the human genome. These modular recombinases can be reprogrammed: New combinations of zinc finger domains and serine recom ...[more]

PMID: 19282480

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