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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The Drosophila Trpm channel mediates calcium influx during egg activation.
Qinan Hu,Mariana F. Wolfner +1 more
TL;DR: The Transient Receptor Potential (TRP) channel subfamily M (Trpm) is identified as a critical channel that mediates the calcium influx and initiated the calcium wave during Drosophila egg activation.
Journal ArticleDOI
Toward establishing an efficient and versatile gene targeting system in higher plants
Masaki Endo,Seiichi Toki +1 more
TL;DR: Recent developments in gene targeting via homologous recombination (GT) are reviewed, giving examples of new techniques improving the efficiency of GT in plants.
Journal ArticleDOI
Large-Scale Transgenic Drosophila Resource Collections for Loss- and Gain-of-Function Studies.
Jonathan Zirin,Yanhui Hu,Lu-Ping Liu,Donghui Yang-Zhou,Ryan Colbeth,Dong Yan,Benjamin Scott Ewen-Campen,Rong Tao,Eric Vogt,Sara VanNest,Cooper Cavers,Christians Villalta,Aram Comjean,Jin Sun,Xia Wang,Yu Jia,Ruibao Zhu,Ping Peng,Jinchao Yu,Da Shen,Yuhao Qiu,Limmond Ayisi,Henna Ragoowansi,Ethan Fenton,Senait Efrem,Annette L. Parks,Kuniaki Saito,Shu Kondo,Liz Perkins,Stephanie E. Mohr,Jian-Quan Ni,Norbert Perrimon,Norbert Perrimon +32 more
TL;DR: The Transgenic RNAi Project (TRiP), a Drosophila melanogaster functional genomics platform at Harvard Medical School, was initiated in 2008 to generate and distribute a genome-scale collection of RNA interference (RNAi) fly stocks, and this work presents an update on these libraries of publicly available RNAi and CRISPR fly stocks.
Journal ArticleDOI
Separate Polycomb Response Elements control chromatin state and activation of the vestigial gene.
Kami Ahmad,Amy E. Spens +1 more
TL;DR: It is shown here that PREs near the wing selector gene vestigial have distinctive roles at their endogenous locus, even though both PREs are repressors in transgenes.
Journal ArticleDOI
An undergraduate laboratory class using CRISPR/Cas9 technology to mutate drosophila genes
Vanesa Adame,Holly Chapapas,Marilyn Cisneros,Carol Deaton,Sophia Deichmann,Chauncey R. Gadek,TyAnna L. Lovato,Maria B. Chechenova,Paul Guerin,Richard M. Cripps +9 more
TL;DR: It is concluded that it is feasible to develop a laboratory genome editing class, to provide effective laboratory training to undergraduate students, and to generate mutant lines for use by the broader scientific community.
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.