Highly efficient genome modifications mediated by CRISPR/Cas9 in Drosophila.
TLDR
The Cas9/gRNA system is an attractive tool for rapid disruption of essentially any gene in Drosophila through targeting seven loci and achieving germline efficiency of up to 100%.Abstract:
We report that Cas9/gRNA mediates efficient genetic modifications in Drosophila. Through targeting seven loci, we achieved a germline efficiency of up to 100%. Genes in both heterochromatin and euchromatin can be modified efficiently. Thus the Cas9/gRNA system is an attractive tool for rapid disruption of essentially any gene in Drosophila.read more
Citations
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CRISPR-Cas systems for editing, regulating and targeting genomes
Jeffry D. Sander,J. Keith Joung +1 more
TL;DR: A modified version of the CRISPR-Cas9 system has been developed to recruit heterologous domains that can regulate endogenous gene expression or label specific genomic loci in living cells, which will undoubtedly transform biological research and spur the development of novel molecular therapeutics for human disease.
Journal ArticleDOI
Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients
Gerald Schwank,Bon-Kyoung Koo,Valentina Sasselli,Johanna F. Dekkers,Inha Heo,Turan Demircan,Nobuo Sasaki,Sander Boymans,Edwin Cuppen,Cornelis K. van der Ent,Edward E. S. Nieuwenhuis,Jeffrey M. Beekman,Hans Clevers +12 more
TL;DR: The CRISPR/Cas9 genome editing system is used to correct the CFTR locus by homologous recombination in cultured intestinal stem cells of CF patients and the corrected allele is expressed and fully functional as measured in clonally expanded organoids.
Journal ArticleDOI
Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila.
TL;DR: A toolbox for high-efficiency genome engineering of Drosophila melanogaster consisting of transgenic Cas9 lines and versatile guide RNA (gRNA) expression plasmids is reported, which will facilitate the rapid evaluation of mutant phenotypes of specific genes and the precise modification of the genome with single-nucleotide precision.
Journal ArticleDOI
Highly Specific and Efficient CRISPR/Cas9-Catalyzed Homology-Directed Repair in Drosophila
Scott J. Gratz,Fiona P. Ukken,C. Dustin Rubinstein,Gene H Thiede,Laura K. Donohue,Alexander M. Cummings,Kate M. O'Connor-Giles +6 more
TL;DR: In this paper, homology-directed repair (HDR) with double-stranded DNA (dsDNA) donor templates was used to enable complex genome engineering through the precise incorporation of large DNA sequences, including screenable markers.
Journal ArticleDOI
Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System
TL;DR: A targeted, continual multigene editing strategy that was applied to the Escherichia coli genome by using the Streptococcus pyogenes type II CRISPR-Cas9 system to realize a variety of precise genome modifications, including gene deletion and insertion, with the highest efficiency of 100%, is described.
References
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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
Efficient genome editing in zebrafish using a CRISPR-Cas system
Woong Y. Hwang,Yanfang Fu,Deepak Reyon,Morgan L. Maeder,Shengdar Q. Tsai,Jeffry D. Sander,Randall T. Peterson,Randall T. Peterson,Jing-Ruey J. Yeh,J. Keith Joung +9 more
TL;DR: It is shown that the CRISPR-Cas system functions in vivo to induce targeted genetic modifications in zebrafish embryos with efficiencies similar to those obtained using zinc finger nucleases and transcription activator-like effector nucleases.
Journal ArticleDOI
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
TL;DR: 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.