Abstract
Serine integrases catalyze precise rearrangement of DNA through site-specific recombination of small sequences of DNA called attachment (att) sites. Unlike other site-specific recombinases, the recombination reaction driven by serine integrases is highly directional and can only be reversed in the presence of an accessory protein called a recombination directionality factor (RDF). The ability to control reaction directionality has led to the development of serine integrases as tools for controlled rearrangement and modification of DNA in synthetic biology, gene therapy, and biotechnology. This review discusses recent advances in serine integrase technologies focusing on their applications in genome engineering, DNA assembly, and logic and data storage devices.
Highlights
Serine integrases catalyze precise rearrangement of DNA through site-specific recombination of small sequences of DNA called attachment sites
The principles of homologous recombination have been applied to DNA assembly[12] and to targeting specific changes in genomic DNA (for example, in nuclease-based genome editing with clustered regularly interspaced short palindromic repeat (CRISPR)-associated nucleases like Cas[9] and Cpf1).[3−8] another group of DNA modifying enzymes the sitespecific recombinases are becoming established as vital tools for genetic modification both in vivo and in vitro
In comparison to tyrosine recombinases, recombination by serine integrases has the advantage of simplicity, normally only requiring the integrase protein and small att sites (
Summary
Sequences of attP and attB are distinct from each other, and from attL and attR sites. The group used a bioinformatics approach to identify a suitable locus for transgene expression, and homologous recombination to insert a target sequence containing genes for antibiotic resistance and GFP flanked by attP sites for phiC31 and Bxb[1] integrases Using these enzymes they integrated transgenic DNA flanked with complementary attB sites, and were able to rapidly generate libraries of embryonic stem cells and induced pluripotent stem cells carrying different combinations of neuronal transcription factor genes. Serine integrases, and their cognate RDFs and att sites, provide reliable DNA recombination systems that can be used in a number of applications including in vivo genomic integration of DNA in a range of organisms, and in vitro assembly and targeted modification of DNA constructs These recombination systems have been applied in low-burden, heritable DNA logic and memory devices that can store information permanently or in a rewritable fashion.
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