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CRISPR/Cas-mediated in planta gene targeting: current advances and challenges

Journal

JOURNAL OF EXPERIMENTAL BOTANY
Volume 74, Issue 13, Pages 3806-3820

Publisher

OXFORD UNIV PRESS
DOI: 10.1093/jxb/erad072

Keywords

CRISPR-Cas; DNA sequence knock-in; DNA sequence replacement; homologous recombination; plant gene targeting

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This paper reviews recent advances in CRISPR/Cas-mediated gene targeting in plants, discusses their potential applications, and suggests further improvements to achieve higher efficiency.
A review of recent advances in CRISPR/Cas-mediated gene targeting in plants, discussing their potential applications, and suggesting further improvements for achieving higher efficiency. Gene targeting can be used to make modifications at a specific region in a plant's genome and create high-precision tools for plant biotechnology and breeding. However, its low efficiency is a major barrier to its use in plants. The discovery of CRISPR (clustered regularly interspaced short palindromic repeats)-Cas-based site-specific nucleases capable of inducing double-strand breaks in desired loci resulted in the development of novel approaches for plant gene targeting. Several studies have recently demonstrated improvements in gene targeting efficiency through cell-type-specific expression of Cas nucleases, the use of self-amplified gene-targeting-vector DNA, or manipulation of RNA silencing and DNA repair pathways. In this review, we summarize recent advances in CRISPR/Cas-mediated gene targeting in plants and discuss potential efficiency improvements. Increasing the efficiency of gene targeting technology will help pave the way for increased crop yields and food safety in environmentally friendly agriculture.

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