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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Genetic knockdown and knockout approaches in Hydra
TL;DR: A CRISPR-Cas9 based targeted mutation approach as well as an optimized, reproducible strategy for the delivery of siRNAs are reported, demonstrating that these strategies provide reliable genetic interference with target gene expression, facilitating functional studies and genome editing in Hydra.
Journal ArticleDOI
Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system
Qingfen Zheng,Li Bai,Sujun Zheng,Mei Liu,J Zhang,Ting Wang,Zhongwei Xu,Yu Chen,Jian-Sheng Li,Zhongping Duan +9 more
TL;DR: The present study aimed to demonstrate the targeted inhibition of DHBV DNA, especially cccDNA, by the CRISPR/Cas9 system using this model.
Journal ArticleDOI
Versatile High-Throughput Fluorescence Assay for Monitoring Cas9 Activity.
TL;DR: A fluorescence assay for monitoring Cas9 nuclease activity and demonstrating its utility with S. pyogenes, S.aureus, and C. jejuni Cas9 is developed and validated, making it broadly applicable for the discovery of additional Cas9 inhibitors or the characterization of Cas9 enzyme variants.
Patent
Manipulation of immunoglobulin gene diversity and multi-antibody therapeutics
TL;DR: In this article, the authors provided improved non-human vertebrates and non-vertebrate cells capable of expressing antibodies comprising human variable region sequences (HCDR3s) comprising variable gene usage common to antibodies useful for treating or preventing certain diseases or conditions.
Journal ArticleDOI
Ret and Substrate-Derived TGF-β Maverick Regulate Space-Filling Dendrite Growth in Drosophila Sensory Neurons.
Nina Hoyer,Philip Zielke,Chun Hu,Meike Petersen,Kathrin Sauter,Robin Scharrenberg,Yun Peng,Charles C. Kim,Chun Han,Jay Z. Parrish,Peter Soba +10 more
TL;DR: The results suggest that Mav acts as a substrate-derived secreted signal promoting dendrite growth within not-yet-covered areas of the receptive field to ensure space-filling dendritic growth.
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.