Permanent Genome Modifications in Plant Cells by Transient Viral Vectors

Genome modifications in plant cells by custom‐made restriction enzymes

“…The stability of the mutation was confirmed by its transmission to the next generation, as confirmed by GUS expression (Figure 4) and DNA sequencing (Marton et al, 2010). As TRV can infect a large number of plant species (Marton et al, 2010;Vainstein et al, 2011) and can travel into a wide variety of cells, tissues and organs, it can potentially be adapted for non-transgenic genome modification in various model and crop plants (Vainstein et al, 2011). Furthermore, because TRV can also travel into existing growing buds, mutations at meristems of growing and developing plants may produce mutated seedlings, similar to the production of mutated seedlings from transgenic Arabidopsis plants (Lloyd et al, 2005;Osakabe et al, 2010;Zhang et al, 2010).…”

“…While relying on the plant's natural or modulated DNA repair mechanisms and using negative-positive selection vectors have proven useful for the targeting of various native and transgenic genes (Table 1), such systems are still challenging and may be difficult to reproduce for the targeting of other genes and in other plant species (reviewed in Hohn and Puchta, 1999;Kumar and Fladung, 2001;Puchta, 2002Puchta, , 2003Britt and May, 2003;Iida and Terada, 2005;Puchta and Hohn, 2005;Porteus, 2009;Weinthal et al, 2010;Vainstein et al, 2011). In the last decade, several advances have been made in the development of NREs and in their deployment for induction of genomic DSBs in eukaryotic cells.…”

“…The plants were then infected with a ZFN-expressing Tobacco rattle virus (TRV) vector, and the virus's ability to travel from cell to cell, express the ZFNs and mutate the genome of these cells was monitored by GUS expression (Figure 4) and sequencing of targeting events. This indirect, virus-aided gene expression (VAGE) system (Vainstein et al, 2011) enabled the production of non-transgenic mutated tobacco and petunia plants. The stability of the mutation was confirmed by its transmission to the next generation, as confirmed by GUS expression (Figure 4) and DNA sequencing (Marton et al, 2010).…”

“…Furthermore, because TRV can also travel into existing growing buds, mutations at meristems of growing and developing plants may produce mutated seedlings, similar to the production of mutated seedlings from transgenic Arabidopsis plants (Lloyd et al, 2005;Osakabe et al, 2010;Zhang et al, 2010). The latter might simplify the production of mutated plants by recovering mutated plantlets from infected adult plants without the need for regeneration through tissue culture (Vainstein et al, 2011).…”

“…Gene targeting was developed into a powerful tool for basic cell research in Saccharomyces cerevisiae (Scherer and Davis, 1979), then was latterly adapted for use in other model organisms including certain fungal species, mouse embryonic stem cells, Drosophila and human cell lines (Baribault and Kemler, 1989;Venken and Bellen, 2005;Porteus, 2007;Hall et al, 2009;Laible and Alonso-Gonzalez, 2009;Tenzen et al, 2009). Ever since these developments, scientists have been seeking to develop similar methods for plant species (Puchta, 2002;Hanin and Paszkowski, 2003;Reiss, 2003;Hohn, 2005, 2010;Tzfira and White, 2005;Porteus, 2009;Weinthal et al, 2010;Vainstein et al, 2011). Yet, gene targeting by homologous recombination (HR), i.e.…”