CRISPR ribonucleoprotein-mediated genetic engineering in plants

Yingxiao Zhang; Brian Iaffaldano; Yiping Qi

doi:10.1016/j.xplc.2021.100168

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Plant Commun. 2021 Feb 10;2(2):100168. doi: 10.1016/j.xplc.2021.100168. eCollection 2021 Mar 8.

Authors

Yingxiao Zhang¹, Brian Iaffaldano¹, Yiping Qi^{1

2}

Affiliations

¹ Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
² Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD 20850, USA.

Abstract

CRISPR-derived biotechnologies have revolutionized the genetic engineering field and have been widely applied in basic plant research and crop improvement. Commonly used Agrobacterium- or particle bombardment-mediated transformation approaches for the delivery of plasmid-encoded CRISPR reagents can result in the integration of exogenous recombinant DNA and potential off-target mutagenesis. Editing efficiency is also highly dependent on the design of the expression cassette and its genomic insertion site. Genetic engineering using CRISPR ribonucleoproteins (RNPs) has become an attractive approach with many advantages: DNA/transgene-free editing, minimal off-target effects, and reduced toxicity due to the rapid degradation of RNPs and the ability to titrate their dosage while maintaining high editing efficiency. Although RNP-mediated genetic engineering has been demonstrated in many plant species, its editing efficiency remains modest, and its application in many species is limited by difficulties in plant regeneration and selection. In this review, we summarize current developments and challenges in RNP-mediated genetic engineering of plants and provide future research directions to broaden the use of this technology.

Keywords: CRISPR; RNP; genetic engineering; genome editing; transgene free.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

MeSH terms

Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
Gene Editing / methods*
Genome, Plant*
Plants / genetics*
Ribonucleoproteins / genetics*

Substances

Ribonucleoproteins