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.read more
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Plant salt-tolerance mechanisms
Ulrich Deinlein,Aaron B. Stephan,Tomoaki Horie,Wei Luo,Wei Luo,Guohua Xu,Julian I. Schroeder +6 more
TL;DR: The understanding of the core salt-tolerance mechanisms in plants is reviewed and key Na+ transport and detoxification pathways and the impact of epigenetic chromatin modifications on salinity tolerance are reviewed.
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A CRISPR/Cas9 toolkit for multiplex genome editing in plants
Hui Li Xing,Li Dong,Zhi-Ping Wang,Hai-Yan Zhang,Chun Yan Han,Bing Liu,Xue Chen Wang,Qi-Jun Chen +7 more
TL;DR: A toolkit that facilitates transient or stable expression of the CRISPR/Cas9 system in a variety of plant species, which will facilitate plant research, as it enables high efficiency generation of mutants bearing multiple gene mutations.
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Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation.
TL;DR: Comparisons of 12 combinations of eight promoters and two terminators found that the efficiency of the egg cell-specific promoter-controlled CRISPR/Cas9 system depended on the presence of a suitable terminator, and the composite promoter generated by fusing two eggcell-specific promoters resulted in much higher efficiency of mutation in the T1 generation compared with the single promoters.
Journal ArticleDOI
Engineering Quantitative Trait Variation for Crop Improvement by Genome Editing.
Daniel Rodríguez-Leal,Zachary H. Lemmon,Jarrett Man,Madelaine E. Bartlett,Zachary B. Lippman +4 more
TL;DR: It is demonstrated that CRISPR/Cas9 genome editing of promoters generates diverse cis-regulatory alleles that provide beneficial quantitative variation for breeding that provide a foundation for dissecting complex relationships between gene-reg regulatory changes and control of quantitative traits.
Journal ArticleDOI
Efficient gene editing in tomato in the first generation using the clustered regularly interspaced short palindromic repeats/CRISPR-associated9 system.
TL;DR: The CRISPR/Cas9 system is highly efficient at generating targeted mutations in stable transgenic tomato plants, and homozygous deletions of a desired size can be created in the first generation.
References
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TL;DR: How NBGT may offer a new perspective for genetic modification is discussed, some aspects crucial for a safety improvement of the corresponding techniques are addressed and the use of NBGT applications and products to the regulatory oversight is related.
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Staying on target with CRISPR-Cas
TL;DR: Four independent studies shed light on the specificity of RNA-guided genome editing tools based on the Streptococcus pyogenes Cas9 protein.
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RNA guides genome engineering.
Claudio Mussolino,Toni Cathomen +1 more
TL;DR: The Cas9 endonuclease is reprogrammed by RNAs for site-specific modification of eukaryotic and microbial genomes.
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Renegotiating GM crop regulation. Targeted gene-modification technology raises new issues for the oversight of genetically modified crops.
Jennifer Kuzma,Adam Kokotovich +1 more
TL;DR: New technologies enable more precise and subtler modification of plant genomes that allow scientists to insert foreign DNA into the plant genome at precise locations, remove unwanted DNA sequences or introduce subtle modifications, such as single‐base substitutions that alter the activity of individual genes.