Geminiviruses for biotechnology: the art of parasite taming

Abstract

58I.58II.59III.59IV.60V.6263References63 Summary Viruses are intracellular pathogens that have evolved efficient strategies for replication and expression of their proteins in the host cells. Geminiviruses – plant viruses with small circular single‐stranded DNA genomes – effectively manipulate plant cell processes for viral functions, entailing great potential for biotechnological applications. This potentiality has been realized in the form of protein expression and gene‐silencing vectors, and, more recently, vectors for genome editing – a technology that these viruses seem particularly well‐suited to facilitate. This insight offers an overview of the biological properties of geminiviruses, with emphasis on those leveraging development of geminivirus‐based replicons. It illustrates the basis for engineering geminivirus‐based replicons and their applications. Furthermore, it discusses the reported use and future perspectives of geminivirus‐based replicons for genome editing.

Improving CRISPR‐Cas‐mediated RNA targeting and gene editing using SPLCV replicon‐based expression vectors in Nicotiana benthamiana

“…Geminiviruses are a family of plant viruses with circular single‐stranded DNA genomes. They have been deconstructed by researchers for multiple biotechnological applications, including protein expression, gene silencing and genome editing, in plants (Lozano‐Durán, 2016). Under the deconstructed virus strategy, the coat protein and movement protein genes were removed from the geminiviruses, while the sequences required for replication were retained.…”

“…Under the deconstructed virus strategy, the coat protein and movement protein genes were removed from the geminiviruses, while the sequences required for replication were retained. The replicons replicate after delivery to plant cells and increase the copy number of carried DNA; this leads to high levels of gene expression (Lozano‐Durán, 2016). Although a few geminiviral replicon‐based vectors have been used in gene targeting (Baltes et al , 2014; Cermak et al , 2015; Wang et al , 2017), the list of DNA replicon‐based vectors is still limited in plants, especially for food crops.…”

Viral Vectors for Plant Genome Engineering

“…Geminiviruses have been engineered as vectors for the expression of heterologous proteins in plants (Lozano-Duran, 2016). Whereas the cargo capacity of these viruses is quite restricted, they can be converted into non-infectious replicons by replacing genes important for infection and cell-to-cell movement with heterologous sequences, such as SSN expression cassettes and repair templates.…”

Barcoding of Plant Viruses with Circular Single-Stranded DNA Based on Rolling Circle Amplification

“…Many more excellent publications have appeared in this context, mainly in the field of classification and taxonomy of known and newly detected ssDNA viruses, as well as of resistance breeding and biotechnology, which cannot be cited here due to space limitations. The reader is referred to comprehensive overviews for those aspects [ 161 , 162 , 163 , 164 , 165 , 166 , 167 , 168 , 169 , 170 ]. The following review rather tries to focus on certain aspects of RCA diagnostics to describe the state-of-the-art technique and its unique advantages.…”