Experts Develop All-in-one CRISPR Toolbox for Large-scale Screens in Plants

Researchers from the University of Maryland, USA, developed a comprehensive all-in-one CRISPR toolbox that addresses the challenge of large-scale CRISPR screens in plants. Their study is published in The Plant Cell.

The new CRISPR toolbox can be effectively used for cytosine base editing (CBE), adenine base editing (ABE), Cas12a-based genome editing and ABE, and CRISPR-Act3.0-based gene activation in both monocot (rice) and dicot (tomato) plants.

To further test how well the vectors work for large-scale gene editing experiments, the scientists created collections of instruction guides called sgRNAs to test in rice cells. This allowed them to quickly identify the most effective guides. They also showed that they could use a library of these guides to make targeted changes in a specific gene in rice, called ACETOLACTATE SYNTHASE. This resulted in finding new versions of the gene that could make the rice resistant to herbicides.

Furthermore, they used the gene activation tool in Arabidopsis, to screen for the best guides to turn on a gene called FLOWERING LOCUS T. Turning this gene on caused the plants to flower much earlier.

This complete set of 61 versatile all-in-one vectors covers almost all the commonly used CRISPR techniques. It will make it much easier to perform large-scale studies to either turn genes off (loss-of-function) or turn them on (gain-of-function) in plants, opening up many exciting possibilities for improving crops and understanding plant biology.

Read the journal article in The Plant Cell.