RNA interference (RNAi) is an evolutionarily conserved, homology
dependent gene silencing mechanism; an area of upfront basic
research, further gaining importance in applications like
development of virus resistant transgenics. To explore the
possibility of transgenic resistance to the Tomato leaf curl virus,
a constraint in tomato production, sub-region of viral replicase
gene (truncated rep) was chosen as target for RNA based silencing.
In silico analysis of rep gene with siRNA design algorithms and
appraisal of rep mRNA, disclosed potent siRNAs and the
accessibility of mRNA for gene silencing. Four different RNAi
constructs viz., antisense T-rep and three hairpin RNA constructs,
were developed and tomato transformed to obtain transformants.
Transformation and regeneration of ToLCV infected leaves resulted
in regeneration of recovered plants. Interestingly, tomato
transformants showed phenotypic anomalies, attributed to the
off-target silencing by siRNAs. Thus the book caters to the plant
molecular biologists and virologists working on the arena of gene
silencing with information on basic criteria for target region
selection, construct design strategies and silencing studies.
General
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