Site- and strand-specific nicking of DNA by fusion proteins derived from MutH and I-SceI or TALE repeats
dc.contributor.author | Gabsalilow, Lilia | |
dc.contributor.author | Schierling, Benno | |
dc.contributor.author | Friedhoff, Peter | |
dc.contributor.author | Pingoud, Alfred | |
dc.contributor.author | Wende, Wolfgang | |
dc.date.accessioned | 2022-11-18T09:57:02Z | |
dc.date.available | 2013-03-04T09:53:47Z | |
dc.date.available | 2022-11-18T09:57:02Z | |
dc.date.issued | 2013 | |
dc.description.abstract | Targeted genome engineering requires nucleases that introduce a highly specific double-strand break in the genome that is either processed by homology-directed repair in the presence of a homologous repair template or by non-homologous end-joining (NHEJ) that usually results in insertions or deletions. The error-prone NHEJ can be efficiently suppressed by nickases that produce a single-strand break rather than a double-strand break. Highly specific nickases have been produced by engineering of homing endonucleases and more recently by modifying zinc finger nucleases (ZFNs) composed of a zinc finger array and the catalytic domain of the restriction endonuclease FokI. These ZF-nickases work as heterodimers in which one subunit has a catalytically inactive FokI domain. We present two different approaches to engineer highly specific nickases; both rely on the sequence-specific nicking activity of the DNA mismatch repair endonuclease MutH which we fused to a DNA-binding module, either a catalytically inactive variant of the homing endonuclease I-SceI or the DNA-binding domain of the TALE protein AvrBs4. The fusion proteins nick strand specifically a bipartite recognition sequence consisting of the MutH and the I-SceI or TALE recognition sequences, respectively, with a more than 1000-fold preference over a stand-alone MutH site. TALE MutH is a programmable nickase. | en |
dc.identifier.uri | http://nbn-resolving.de/urn:nbn:de:hebis:26-opus-92283 | |
dc.identifier.uri | https://jlupub.ub.uni-giessen.de//handle/jlupub/9668 | |
dc.identifier.uri | http://dx.doi.org/10.22029/jlupub-9056 | |
dc.language.iso | en | de_DE |
dc.rights | Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 International | * |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/3.0/ | |
dc.subject.ddc | ddc:570 | de_DE |
dc.title | Site- and strand-specific nicking of DNA by fusion proteins derived from MutH and I-SceI or TALE repeats | en |
dc.type | article | de_DE |
local.affiliation | FB 08 - Biologie und Chemie | de_DE |
local.opus.fachgebiet | Biochemie (FB 08) | de_DE |
local.opus.id | 9228 | |
local.opus.institute | Institute for Biochemistry | de_DE |
local.source.freetext | Nucleic Acids Research; | de_DE |
local.source.uri | https://doi.org/10.1093/nar/gkt080 |
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