REAL

Conditional DNA repair mutants enable highly precise genome engineering

Nyerges, Ákos and Csorgő, Bálint and Nagy, István and Latinovics, Dóra and Szamecz, Béla and Pósfai, György and Pál, Csaba (2014) Conditional DNA repair mutants enable highly precise genome engineering. NUCLEIC ACIDS RESEARCH, 42 (8). e62. ISSN 0305-1048

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Abstract

Oligonucleotide-mediated multiplex genome engineering is an important tool for bacterial genome editing. The efficient application of this technique requires the inactivation of the endogenous methyl-directed mismatch repair system that in turn leads to a drastically elevated genomic mutation rate and the consequent accumulation of undesired off-target mutations. Here, we present a novel strategy for mismatch repair evasion using temperature-sensitive DNA repair mutants and temporal inactivation of the mismatch repair protein complex in Escherichia coli. Our method relies on the transient suppression of DNA repair during mismatch carrying oligonucleotide integration. Using temperature-sensitive control of methyl-directed mismatch repair protein activity during multiplex genome engineering, we reduced the number of off-target mutations by 85%, concurrently maintaining highly efficient and unbiased allelic replacement.

Item Type: Article
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia > QH3015 Molecular biology / molekuláris biológia
Q Science / természettudomány > QH Natural history / természetrajz > QH426 Genetics / genetika, örökléstan
Depositing User: Dr. Béla Szamecz
Date Deposited: 25 Sep 2014 09:55
Last Modified: 04 Oct 2014 01:14
URI: http://real.mtak.hu/id/eprint/16693

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