13 research outputs found
In situ functional dissection of RNA cis-regulatory elements by multiplex CRISPR-Cas9 genome engineering
RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene
regulation. Deciphering the prevalence and functional impact of this post-transcriptional control
layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we
describe genome-engineering based evaluation of RNA regulatory element activity (GenERA), a
CRISPR-Cas9 platform for in situ high-content functional analysis of RREs. We use GenERA to
survey the entire regulatory landscape of a 3âUTR, and apply it in a multiplex fashion to analyse
combinatorial interactions between sets of miRNA response elements (MREs), providing strong
evidence for cooperative activity. We also employ this technology to probe the functionality of an
entire MRE network under cellular homeostasis, and show that high-resolution analysis of the
GenERA dataset can be used to extract functional features of MREs. This study provides a
genome editing-based multiplex strategy for direct functional interrogation of RNA cis-regulatory
elements in a native cellular environment
Genomic Designing for Climate-Smart Tomato
Tomato is the first vegetable consumed in the world. It is grown in very different conditions and areas, mainly in field for processing tomatoes while fresh-market tomatoes are often produced in greenhouses. Tomato faces many environmental stresses, both biotic and abiotic. Today many new genomic resources are available allowing an acceleration of the genetic progress. In this chapter, we will first present the main challenges to breed climate-smart tomatoes. The breeding objectives relative to productivity, fruit quality, and adaptation to environmental stresses will be presented with a special focus on how climate change is impacting these objectives. In the second part, the genetic and genomic resources available will be presented. Then, traditional and molecular breeding techniques will be discussed. A special focus will then be presented on ecophysiological modeling, which could constitute an important strategy to define new ideotypes adapted to breeding objectives. Finally, we will illustrate how new biotechnological tools are implemented and could be used to breed climate-smart tomatoes