Oligo-Mediated Targeted Gene Editing

Gocal, Greg F. W.; Schöpke, C.; Beetham, Peter R.

doi:10.1007/978-1-4939-2556-8_5

Cited by 26 publications

(31 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These modifications prevented the oligonucleotides from undergoing recombination, but still allowed them to act as a mutagen and DNA template. For this reason, ODM is considered a targeted mutagenesis system (Dong et al, 2006;Gocal et al, 2015). Based on peer-reviewed literature, the most frequently used designs were modified with phosphorothioate linkages on the terminal bases ( Figure 1b; Andrieu-Soler et al, 2005;De Pi edoue et al, 2007;Radecke et al, 2006).…”

Section: The Oligonucleotide Templatementioning

confidence: 99%

“…These alterations may also include precise insertions or deletions in the target sequence. The oligonucleotide-directed mutagenesis (ODM) technique for genome editing has been successfully employed in bacterial, yeast, mammalian and plant systems Gocal et al, 2015;Moerschell et al, 1988;Yoon et al, 1996). This review will specifically focus on using chemically synthesized oligonucleotides as a template for making targeted changes in these four systems.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Oligonucleotide‐directed mutagenesis for precision gene editing

Sauer

Mozoruk

Miller

et al. 2015

Plant Biotechnology Journal

Self Cite

129

View full text Add to dashboard Cite

SummaryDifferences in gene sequences, many of which are single nucleotide polymorphisms, underlie some of the most important traits in plants. With humanity facing significant challenges to increase global agricultural productivity, there is an urgent need to accelerate the development of these traits in plants. oligonucleotide‐directed mutagenesis (ODM), one of the many tools of Cibus’ Rapid Trait Development System (RTDS ™) technology, offers a rapid, precise and non‐transgenic breeding alternative for trait improvement in agriculture to address this urgent need. This review explores the application of ODM as a precision genome editing technology, with emphasis on using oligonucleotides to make targeted edits in plasmid, episomal and chromosomal DNA of bacterial, fungal, mammalian and plant systems. The process of employing ODM by way of RTDS technology has been improved in many ways by utilizing a fluorescence conversion system wherein a blue fluorescent protein (BFP) can be changed to a green fluorescent protein (GFP) by editing a single nucleotide of the BFP gene (CAC→TAC; H66 to Y66). For example, dependent on oligonucleotide length, applying oligonucleotide‐mediated technology to target the BFP transgene in Arabidopsis thaliana protoplasts resulted in up to 0.05% precisely edited GFP loci. Here, the development of traits in commercially relevant plant varieties to improve crop performance by genome editing technologies such as ODM, and by extension RTDS, is reviewed.

show abstract

Section: The Oligonucleotide Templatementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Oligonucleotide‐directed mutagenesis for precision gene editing

Sauer

Mozoruk

Miller

et al. 2015

Plant Biotechnology Journal

Self Cite

129

View full text Add to dashboard Cite

show abstract

“…Once the correction process is completed, the oligos are degraded. In in vitro assays using "cell-free" extracts and various pure enzymes, it was demonstrated that the oligos are stable for sufficient time to direct gene correction and then are quickly degraded (Gocal et al 2015).…”

Section: Methods Description and List Of Techniquesmentioning

confidence: 99%

New developments in green biotechnology - an inventory for RIVM

Wiel¹,

Smulders

Visser³

et al. 2016

View full text Add to dashboard Cite

“…Novel methods for targeted editing of plant genomic DNA will play an important role in addressing these challenges. Oligonucleotide-directed mutagenesis (ODM) is a nontransgenic base pair-specific precision genome editing platform that has been employed successfully in bacterial, fungal, mammalian, and plant systems (Moerschell et al, 1988;Yoon et al, 1996;Beetham et al, 1999;Zhu et al, 1999;Aarts et al, 2006;Gocal et al, 2015;Sauer et al, 2016). ODM employs chemically synthesized oligonucleotides to mediate genome editing by acting as DNA templates during the editing process.…”

mentioning

confidence: 99%

“…ODM employs chemically synthesized oligonucleotides to mediate genome editing by acting as DNA templates during the editing process. In the early application of this technology in plants, oligonucleotides were used to target the acetolactate synthase (ALS) gene in tobacco (Nicotiana tabacum; Beetham et al, 1999), in maize (Zea mays; Zhu et al, 1999), and later in oilseed rape (Brassica napus; Gocal et al, 2015). In both tobacco and maize, a DNA/RNA chimeric oligonucleotide was used to target the ALS gene, whereas for oilseed rape, an oligonucleotide containing a 59Cy3 dye and a 39idC reverse base was employed.…”

mentioning

confidence: 99%

Oligonucleotide-Mediated Genome Editing Provides Precision and Function to Engineered Nucleases and Antibiotics in Plants

et al. 2016

Self Cite

View full text Add to dashboard Cite

Here, we report a form of oligonucleotide-directed mutagenesis for precision genome editing in plants that uses single-stranded oligonucleotides (ssODNs) to precisely and efficiently generate genome edits at DNA strand lesions made by DNA double strand break reagents. Employing a transgene model in Arabidopsis (Arabidopsis thaliana), we obtained a high frequency of precise targeted genome edits when ssODNs were introduced into protoplasts that were pretreated with the glycopeptide antibiotic phleomycin, a nonspecific DNA double strand breaker. Simultaneous delivery of ssODN and a site-specific DNA double strand breaker, either transcription activator-like effector nucleases (TALENs) or clustered, regularly interspaced, short palindromic repeats (CRISPR/Cas9), resulted in a much greater targeted genome-editing frequency compared with treatment with DNA double strand-breaking reagents alone. Using this site-specific approach, we applied the combination of ssODN and CRISPR/Cas9 to develop an herbicide tolerance trait in flax (Linum usitatissimum) by precisely editing the 59-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE (EPSPS) genes. EPSPS edits occurred at sufficient frequency that we could regenerate whole plants from edited protoplasts without employing selection. These plants were subsequently determined to be tolerant to the herbicide glyphosate in greenhouse spray tests. Progeny (C1) of these plants showed the expected Mendelian segregation of EPSPS edits. Our findings show the enormous potential of using a genome-editing platform for precise, reliable trait development in crop plants.

show abstract

Oligo-Mediated Targeted Gene Editing

Cited by 26 publications

References 34 publications

Oligonucleotide‐directed mutagenesis for precision gene editing

Oligonucleotide‐directed mutagenesis for precision gene editing

New developments in green biotechnology - an inventory for RIVM

Oligonucleotide-Mediated Genome Editing Provides Precision and Function to Engineered Nucleases and Antibiotics in Plants

Contact Info

Product

Resources

About