Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice
Reads0
Chats0
TLDR
Adaptations of the type II CRISPR/Cas system leading to successful expression of the Cas9/sgRNA system in model plant and crop species bodes well for its near-term use as a facile and powerful means of plant genetic engineering for scientific and agricultural applications.Abstract:
The type II CRISPR/Cas system from Streptococcus pyogenes and its simplified derivative, the Cas9/single guide RNA (sgRNA) system, have emerged as potent new tools for targeted gene knockout in bacteria, yeast, fruit fly, zebrafish and human cells. Here, we describe adaptations of these systems leading to successful expression of the Cas9/sgRNA system in two dicot plant species, Arabidopsis and tobacco, and two monocot crop species, rice and sorghum. Agrobacterium tumefaciens was used for delivery of genes encoding Cas9, sgRNA and a non-fuctional, mutant green fluorescence protein (GFP) to Arabidopsis and tobacco. The mutant GFP gene contained target sites in its 5' coding regions that were successfully cleaved by a CAS9/sgRNA complex that, along with error-prone DNA repair, resulted in creation of functional GFP genes. DNA sequencing confirmed Cas9/sgRNA-mediated mutagenesis at the target site. Rice protoplast cells transformed with Cas9/sgRNA constructs targeting the promoter region of the bacterial blight susceptibility genes, OsSWEET14 and OsSWEET11, were confirmed by DNA sequencing to contain mutated DNA sequences at the target sites. Successful demonstration of the Cas9/sgRNA system in model plant and crop species bodes well for its near-term use as a facile and powerful means of plant genetic engineering for scientific and agricultural applications.read more
Citations
More filters
Book ChapterDOI
Improvement of Crop’s Stress Tolerance by Gene Editing CRISPR/CAS9 System
Avinash Singh,Rajib Roychowdhury,Toolika Singh,Wenjing Wang,Deepanker Yadav,Ajay Kumar,Arpan Modi,Avinash Chandra Rai,Sandeep Ghughe,Anil Kumar,Prashant Kumar Singh,Prashant Kumar Singh +11 more
TL;DR: CRISPR/Cas9-mediated genome editing (CMGE) has revolutionized agriculture by offering a tool for trait improvement, gene regulation, development of virus resistance, and the generation of mutant libraries.
Book ChapterDOI
The CRISPR/Cas9 System for Crop Improvement: Progress and Prospects
Kah-Yung Bernard Leong,Yee-Han Chan,Wan Muhamad Asrul Nizam Wan Abdullah,Swee Hua Erin Lim,Kok-Song Lai +4 more
TL;DR: This chapter aims to highlight the progress and application of genome editing techniques, in particular, the CRISPR/Cas9 system as a powerful genome editing tool for crop improvement.
Journal ArticleDOI
Genome engineering technologies for targeted genetic modification in plants
Wei Tang,Anna Y. Tang +1 more
TL;DR: A detailed overview of these systems is provided, the strengths and weaknesses of each are highlighted, research advances made with these technologies in model and crop plants are summarized, and their applications in plant functional genomics are discussed.
Posted ContentDOI
CRISPR-Cas9 Mediated Knockout of NtAn1 to Enhance the Lipid Accumulation in Tobacco Seed for Biodiesel Production
TL;DR: The present results revealed that CRISPR-Cas9 system could be employed in tobacco seed de novo domestication for biodiesel feedstock production and regulated NtAn1 genes regulate both PAs and lipid accumulation in the process of seed development.
Investigating HOX Protein Requirement for Tarsus Determination in Drosophila melanogaster
TL;DR: This paper aims to provide a chronology of the events that led to and culminated in the publication of the first book in the series, “On the Road to Serengeti: Foundations of a Journals” (2003).
References
More filters
Journal ArticleDOI
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
Martin Jinek,Krzysztof Chylinski,Krzysztof Chylinski,Ines Fonfara,Michael H. Hauer,Jennifer A. Doudna,Emmanuelle Charpentier +6 more
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
Le Cong,Le Cong,F. Ann Ran,F. Ann Ran,David M. Cox,David M. Cox,Shuailiang Lin,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +15 more
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
Le Cong,F. A. Ran,David Benjamin Turitz Cox,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +10 more
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
Prashant Mali,Luhan Yang,Kevin M. Esvelt,John Aach,Marc Güell,James E. DiCarlo,Julie E. Norville,George M. Church,George M. Church +8 more
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.
Journal ArticleDOI
Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression.
Lei S. Qi,Matthew H. Larson,Luke A. Gilbert,Jennifer A. Doudna,Jonathan S. Weissman,Adam P. Arkin,Adam P. Arkin,Wendell A. Lim +7 more
TL;DR: This RNA-guided DNA recognition platform provides a simple approach for selectively perturbing gene expression on a genome-wide scale and can efficiently repress expression of targeted genes in Escherichia coli, with no detectable off-target effects.