Agroinfiltration Mediated Scalable Transient Gene Expression in Genome Edited Crop Plants.

Maninder Kaur,Kamel A Abd-Elsalam,Kamil Kuca,Anu Kalia,Farah K Ahmed,Pooja Manchanda,Eugenie Nepovimova

doi:10.3390/ijms221910882

Abstract

Agrobacterium-mediated transformation is one of the most commonly used genetic transformation method that involves transfer of foreign genes into target plants. Agroinfiltration, an Agrobacterium-based transient approach and the breakthrough discovery of CRISPR/Cas9 holds trending stature to perform targeted and efficient genome editing (GE). The predominant feature of agroinfiltration is the abolishment of Transfer-DNA (T-DNA) integration event to ensure fewer biosafety and regulatory issues besides showcasing the capability to perform transcription and translation efficiently, hence providing a large picture through pilot-scale experiment via transient approach. The direct delivery of recombinant agrobacteria through this approach carrying CRISPR/Cas cassette to knockout the expression of the target gene in the intercellular tissue spaces by physical or vacuum infiltration can simplify the targeted site modification. This review aims to provide information on Agrobacterium-mediated transformation and implementation of agroinfiltration with GE to widen the horizon of targeted genome editing before a stable genome editing approach. This will ease the screening of numerous functions of genes in different plant species with wider applicability in future.

Highlights

The CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic repeat DNASequences- CRISPR associated), a Nobel prize-winning technology for the year 2020 pioneered by Emmanuelle Charpentier and Jennifer Doudna in 2012 proved to be an indispensable tool for editing genomes in the agricultural biotechnology field
Keeping in view the generation of stable transformation through Agrobacterium-based technique that involves tissue culture, we focus on Agrobacterium-based transient agroinfiltration approach which does not involve time-consuming tissue culturing step [17]
Agroinfiltration coupled with genome editing holds promise for targeted site modification in the genomic sequence without the integration of the construct into the plant genome

Summary

Introduction

Sequences- CRISPR associated), a Nobel prize-winning technology for the year 2020 pioneered by Emmanuelle Charpentier and Jennifer Doudna in 2012 proved to be an indispensable tool for editing genomes in the agricultural biotechnology field. The discoveries of meganucleases, zinc finger nucleases (ZFNs), Transcription Activator-Like Effector Nucleases (TALENs) followed by the breakthrough discovery of CRISPR-Cas have revolutionized the genome editing research. It involves targeted base editing of the genome through the use of Streptococcus pyogenes Cas endonuclease and short guide RNA molecule (sgRNA) that revolutionized genetics and functional genomics [2,3]. DNA double-stranded breaks in the targeted genome sequences and ensure successful editing of plant genomes [4]

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