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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Book ChapterDOI
Drosophila melanogaster Oogenesis: An Overview
TL;DR: A brief introduction to Drosophila oogenesis is provided, along with a survey of its diverse biological topics and the advanced genetic tools that continue to make this a popular developmental model system.
Journal ArticleDOI
CRISPR/Cas9-Directed Genome Editing of Cultured Cells
TL;DR: The protocol for Cas9‐mediated human genome engineering is described, including construct building and transfection methods necessary for delivering Cas9 and guide RNA (gRNA) into human‐induced pluripotent stem cells (hiPSCs) and HEK293 cells.
Journal ArticleDOI
Heritable Genome Editing with CRISPR/Cas9 in the Silkworm, Bombyx mori
TL;DR: The CRISPR/Cas9 system can act as a highly specific and heritable gene-editing tool in Bombyx mori and is proved that genome alterations at specific sites could be induced by direct microinjection of specific guide RNA and Cas9-mRNA into silkworm embryos.
Journal ArticleDOI
An efficient CRISPR-based strategy to insert small and large fragments of DNA using short homology arms.
Oguz Kanca,Oguz Kanca,Jonathan Zirin,Jonathan Zirin,Jorge Garcia-Marques,Shannon Marie Knight,Shannon Marie Knight,Donghui Yang-Zhou,Donghui Yang-Zhou,Gabriel Amador,Gabriel Amador,Hyunglok Chung,Hyunglok Chung,Zhongyuan Zuo,Zhongyuan Zuo,Liwen Ma,Yuchun He,Wen-Wen Lin,Ying Fang,Ming Ge,Shinya Yamamoto,Karen L. Schulze,Karen L. Schulze,Yanhui Hu,Yanhui Hu,Allan C. Spradling,Stephanie E. Mohr,Stephanie E. Mohr,Norbert Perrimon,Norbert Perrimon,Hugo J. Bellen +30 more
TL;DR: Three new simpler ways to edit genes in flies using single stranded DNA donors using PCR and add 100 nt of homology on each side of an integration cassette, followed by enzymatic removal of one strand.
Journal ArticleDOI
Systematic Evaluation of Drosophila CRISPR Tools Reveals Safe and Robust Alternatives to Autonomous Gene Drives in Basic Research.
TL;DR: The method is compared to recently developed autonomous gene drive technology for somatic and germline genome engineering and concludes that optimized CRISPR with independent transgenes is as efficient, more versatile, and does not represent a biosafety risk.
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.