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When more is better: multigene engineering in plants

2009; Elsevier BV; Volume: 15; Issue: 1 Linguagem: Inglês

10.1016/j.tplants.2009.09.010

1878-4372

Shaista Naqvi, Gemma Farré, Georgina Sanahuja, Teresa Capell, Changfu Zhu, Paul Christou,

CRISPR and Genetic Engineering

The genomics revolution has taught us that a great deal of information can be derived from studying many genes or proteins at the same time. We are beginning to see this approach blossoming in applied research. Instead of attempting to generate useful transgenic plants by introducing single genes, we now see an increasing number of researchers embracing multigene transfer (MGT) as an approach to generate plants with more ambitious phenotypes. MGT allows researchers to achieve goals that were once impossible – the import of entire metabolic pathways, the expression of entire protein complexes, the development of transgenic crops simultaneously engineered to produce a spectrum of added-value compounds. The potential appears limitless. The genomics revolution has taught us that a great deal of information can be derived from studying many genes or proteins at the same time. We are beginning to see this approach blossoming in applied research. Instead of attempting to generate useful transgenic plants by introducing single genes, we now see an increasing number of researchers embracing multigene transfer (MGT) as an approach to generate plants with more ambitious phenotypes. MGT allows researchers to achieve goals that were once impossible – the import of entire metabolic pathways, the expression of entire protein complexes, the development of transgenic crops simultaneously engineered to produce a spectrum of added-value compounds. The potential appears limitless. a binary vector for plant transformation based on the bacterial artificial chromosome (BAC) which is maintained as a large, low copy number vector in E. coli and facilitates the cloning of large DNA fragments. the linking of DNA molecules in tandem to create end-to-end arrays. the simultaneous introduction of two or more transgenes. interruptions in the DNA sugar–phosphate backbone which provide free ends that act as substrates for a variety of repair enzymes. a cloning vector into which DNA fragments are introduced by site-specific recombination rather than the standard restriction and ligation reaction. An entry vector containing the transgene of interest can be used to transfer the transgene to any number of different destination vectors by exploiting site-specific recombination between the bacteriophage λ attR and attL sites which are included in the vector system (LR recombination). recombination between DNA sequences that does not require any homology between the recombining partners nor any specific DNA sequences. Might involve transient pairing between regions of microhomology but also includes homology-free end-to-end joining. transformation with two or more transgenes linked in cis on the same vector or T-DNA. a process for introducing several or many transgenes into a plant simultaneously. a plant vector containing the minimal components to function as a chromosome, specifically DNA sequences comprising a plant centromere and a selectable and/or visible marker. sequence similarity over very short regions (typically <10 bp). a series of genes expressed as a polycistronic mRNA. transgenes that confer a desired phenotype in a transgenic plant, as opposed to a marker gene used for selection. Can be called a structural transgene if the production and extraction of a specific value-added molecule is the goal of the gene transfer experiment. stable multiple arrays of transgenes. A transgenic locus containing multiple genes, which is stable through meiosis so as to avoid segregation, rearrangement and epigenetic silencing. bringing transgenes together in a single plant line through sequential crosses. the transformation of plants which are already transgenic for a different transgene. transformation competent artificial chromosome. A high-capacity plant vector based on the PAC (P1 artificial chromosome) vector of E. coli. an enzyme which relaxes torsional strain in double-stranded DNA molecules by passing one strand through a transient gap in the other. Often found at replication forks and repair complexes. a segment of exogenous DNA containing one or more genes. transformation with two or more transgenes provided on different vectors or T-DNAs.