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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

CRISPR-Local: a local single-guide RNA (sgRNA) design tool for non-reference plant genomes.

TL;DR: CRISPR-Local outperforms other sgRNA design tools in the following respects: designing sg RNAs suitable for non-reference varieties; screening for sgRNAs that are capable of simultaneously targeting multiple genes; saving computational resources by avoiding repeated calculations from multiple submissions; and running offline, with both command-line and graphical user interface modes.
Journal ArticleDOI

CsLFY is required for shoot meristem maintenance via interaction with WUSCHEL in cucumber (Cucumis sativus)

TL;DR: It is suggested that CsLFY has a novel function in regulating shoot meristem maintenance through interaction with CsWUS, and promotes flower development via activation of CsAP3 and CUM1 in cucumber.
Patent

Compositions and methods for producing plants resistant to glyphosate herbicide

TL;DR: In this paper, a guide polynucleotide/Cas endonuclease system was used to identify and insert a double strand break at a specific target site into the genome of a cell, where the nucleotide sequence of interest to be edited can be located within or outside the target site that is recognized by a Cas end-uclease.
Journal ArticleDOI

Hotter, drier, CRISPR: the latest edit on climate change

TL;DR: In this article, the integration of genome editing and transgenics into current breeding strategies is one promising solution to accelerate genetic gains through targeted genetic modifications, producing crops that can overcome the shifting climate realities.
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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