Multiplex genomic structure variation mediated by TALEN and ssODN

Abstract

BackgroundGenomic structure variation (GSV) is widely distributed in various organisms and is an important contributor to human diversity and disease susceptibility. Efficient approaches to induce targeted genomic structure variation are crucial for both analytic and therapeutic studies of GSV. Here, we presented an efficient strategy to induce targeted GSV including chromosomal deletions, duplications and inversions in a precise manner.ResultsUtilizing Transcription Activator-Like Effector Nucleases (TALEN) designed to target two distinct sites, we demonstrated targeted deletions, duplications and inversions of an 8.9 Mb chromosomal segment, which is about one third of the entire chromosome. We developed a novel method by combining TALEN-induced GSV and single stranded oligodeoxynucleotide (ssODN) mediated gene modifications to reduce unwanted mutations occurring during the targeted GSV using TALEN or Zinc finger nuclease (ZFN). Furthermore, we showed that co-introduction of TALEN and ssODN generated unwanted complex structure variation other than the expected chromosomal deletion.ConclusionsWe demonstrated the ability of TALEN to induce targeted GSV and provided an efficient strategy to perform GSV precisely. Furthermore, it is the first time to show that co-introduction of TALEN and ssODN generated unwanted complex structure variation. It is plausible to believe that the strategies developed in this study can be applied to other organisms, and will help understand the biological roles of GSV and therapeutic applications of TALEN and ssODN.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-41) contains supplementary material, which is available to authorized users.

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Genome Editing of Structural Variations: Modeling and Gene Correction

“…This method is more desirable in terms of safety because it can completely prevent nonspecific genomic integration. Successful chromosomal inversions have also been reported in insect and annelid models using customized TALEN systems [36,37]. Nucleases with low DNA specificity should be used with caution because they are likely to have off-target effects that can cause unwanted modification [30,38].…”

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