Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease
Scott J. Gratz,Alexander M. Cummings,Jennifer N. Nguyen,Danielle C. Hamm,Laura K. Donohue,Melissa M. Harrison,Jill Wildonger,Kate M. O'Connor-Giles +7 more
TLDR
A bacterial CRISPR RNA/Cas9 system is adapted to precisely engineer the Drosophila genome and it is reported that Cas9-mediated genomic modifications are efficiently transmitted through the germline.Abstract:
We have adapted a bacterial CRISPR RNA/Cas9 system to precisely engineer the Drosophila genome and report that Cas9-mediated genomic modifications are efficiently transmitted through the germline. This RNA-guided Cas9 system can be rapidly programmed to generate targeted alleles for probing gene function in Drosophila.read more
Citations
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Highly Improved Gene Targeting by Germline-Specific Cas9 Expression in Drosophila
Shu Kondo,Ryu Ueda +1 more
TL;DR: A simple yet extremely efficient platform for systematic gene targeting by the RNA-guided endonuclease Cas9 in Drosophila, which demonstrates rapid generation of mutants in seven neuropeptide and two microRNA genes in which no mutants have been described.
Journal ArticleDOI
Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair
Thomas O. Auer,Karine Duroure,Karine Duroure,Karine Duroure,Anne De Cian,Anne De Cian,Jean-Paul Concordet,Jean-Paul Concordet,Filippo Del Bene,Filippo Del Bene,Filippo Del Bene +10 more
TL;DR: CRISPR/Cas9-mediated knock-in of DNA cassettes into the zebrafish genome at a very high rate by homology-independent double-strand break (DSB) repair pathways is reported and the possibility of easily targeting DNA integration at endogenous loci is shown, thus greatly facilitating the creation of reporter and loss-of-function alleles.
Journal ArticleDOI
CRISPR/Cas9 for genome editing: progress, implications and challenges
Feng Zhang,Yan Wen,Xiong Guo +2 more
TL;DR: The molecular mechanism, applications and challenges of CRISPR/Cas9-mediated genome editing and clinical therapeutic potential ofCRISPR-associated protein 9 are reviewed and a protospacer adjacent motif locating at downstream of target sequences is reviewed.
Journal ArticleDOI
Genome Engineering with Targetable Nucleases
TL;DR: Three classes of targetable cleavage reagents are described: zinc-finger nucleases, transcription activator-like effector nucleases (TALENs), and CRISPR/Cas RNA-guided nuclease (RGNs), which have been successfully used to modify genomic sequences in a wide variety of cells and organisms, including humans.
Journal ArticleDOI
A double-edged sword: R loops as threats to genome integrity and powerful regulators of gene expression
TL;DR: Recent findings about R loops, three-stranded nucleic acid structures that comprise nascent RNA hybridized with the DNA template, leaving the nontemplate DNA single-STRanded, are reviewed.
References
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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
CRISPR provides acquired resistance against viruses in prokaryotes
Rodolphe Barrangou,Christophe Fremaux,Hélène Deveau,Melissa Richards,Patrick Boyaval,Sylvain Moineau,Dennis A. Romero,Philippe Horvath +7 more
TL;DR: It is found that, after viral challenge, bacteria integrated new spacers derived from phage genomic sequences, and CRISPR provided resistance against phages, and resistance specificity is determined by spacer-phage sequence similarity.