Zinc Finger Recombinases with Adaptable DNA Sequence Specificity
| Autor: | Arlene L. McPherson, Christopher Proudfoot, W. Marshall Stark, Andreas F. Kolb |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Protein Conformation
lcsh:Medicine Protein Engineering Biochemistry chemistry.chemical_compound Molecular Cell Biology Recombinase Genomic library lcsh:Science Zinc finger Genetics Recombination Genetic Multidisciplinary Caseins Zinc Fingers Enzymes Nucleic acids Genetic Engineering Research Article Biotechnology Protein Binding DNA recombination Molecular Sequence Data Biology Molecular Genetics Recombinases Genetic Mutation Sequence Homology Nucleic Acid DNA-binding proteins Animals Site-specific recombination Gene Gene Library Recombinase activity Binding Sites Base Sequence lcsh:R Proteins Genetic Variation DNA Zinc finger nuclease Protein Structure Tertiary chemistry Mutagenesis Enzyme Structure Protein structure Biocatalysis lcsh:Q Cattle |
| Zdroj: | PLoS ONE Proudfoot, C, McPherson, A L, Kolb, A F & Stark, W M 2011, ' Zinc finger recombinases with adaptable DNA sequence specificity ', PLoS ONE, vol. 6, no. 4, pp. e19537 . https://doi.org/10.1371/journal.pone.0019537 PLoS ONE, Vol 6, Iss 4, p e19537 (2011) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0019537 |
| Popis: | Site-specific recombinases have become essential tools in genetics and molecular biology for the precise excision or integration of DNA sequences. However, their utility is currently limited to circumstances where the sites recognized by the recombinase enzyme have been introduced into the DNA being manipulated, or natural ‘pseudosites’ are already present. Many new applications would become feasible if recombinase activity could be targeted to chosen sequences in natural genomic DNA. Here we demonstrate efficient site-specific recombination at several sequences taken from a 1.9 kilobasepair locus of biotechnological interest (in the bovine b-casein gene), mediated by zinc finger recombinases (ZFRs), chimaeric enzymes with linked zinc finger (DNA recognition) and recombinase (catalytic) domains. In the "Z-sites" tested here, 22 bp casein gene sequences are flanked by 9 bp motifs recognized by zinc finger domains. Asymmetric Z-sites were recombined by the concomitant action of two ZFRs with different zinc finger DNA-binding specificities, and could be recombined with a heterologous site in the presence of a third recombinase. Our results show that engineered ZFRs may be designed to promote site-specific recombination at many natural DNA sequences. |
| Databáze: | OpenAIRE |
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