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Open AccessJournal ArticleDOI

Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system

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TLDR
The CRISPR/Cas system allows targeted cleavage of genomic DNA guided by a customizable small noncoding RNA, resulting in gene modifications by both non-homologous end joining (NHEJ) and homology-directed repair (HDR) mechanisms.
Abstract
Targeted genome engineering (also known as genome editing) has emerged as an alternative to classical plant breeding and transgenic (GMO) methods to improve crop plants. Until recently, available tools for introducing site-specific double strand DNA breaks were restricted to zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs). However, these technologies have not been widely adopted by the plant research community due to complicated design and laborious assembly of specific DNA binding proteins for each target gene. Recently, an easier method has emerged based on the bacterial type II CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) immune system. The CRISPR/Cas system allows targeted cleavage of genomic DNA guided by a customizable small noncoding RNA, resulting in gene modifications by both non-homologous end joining (NHEJ) and homology-directed repair (HDR) mechanisms. In this review we summarize and discuss recent applications of the CRISPR/Cas technology in plants.

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Citations
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Journal ArticleDOI

Recent Progress in Genetic Transformation and Gene Editing Technology in Cucurbit Crops

TL;DR: In this paper , the authors summarize recent advances in improving the genetic transformation efficiency of cucurbit crops, including the screening of germplasm and the application of physical treatments, morphogenic genes, and selection markers.

Evaluation of reagents and methods for genome editing in potato (Solanum tuberosum L.)

TL;DR: The evaluated reagents and methods provide a frame work for genome editing in potato and other vegetatively propagated crops and have important implications for basic research and agriculture.
Book ChapterDOI

Transformation and T-DNA Mutagenesis

TL;DR: High-efficiency transformation has enabled the creation of a large collection of sequence indexed insertional mutants, and this T-DNA collection is a valuable experimental resource that has been utilized by hundreds of researchers around the world.
References
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Journal ArticleDOI

A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.

TL;DR: This study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
Journal ArticleDOI

Multiplex Genome Engineering Using CRISPR/Cas Systems

TL;DR: The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage as discussed by the authors.

Multiplex Genome Engineering Using CRISPR/Cas Systems

TL;DR: Two different type II CRISPR/Cas systems are engineered and it is demonstrated that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.
Journal ArticleDOI

RNA-Guided Human Genome Engineering via Cas9

TL;DR: The type II bacterial CRISPR system is engineer to function with custom guide RNA (gRNA) in human cells to establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
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