Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish
An Xiao,Zhanxiang Wang,Yingying Hu,Yingdan Wu,Zhou Luo,Zhipeng Yang,Yao Zu,Wenyuan Li,Peng Huang,Xiangjun Tong,Zuoyan Zhu,Shuo Lin,Bo Zhang +12 more
TLDR
These findings indicate that both TALENs and Cas9/gRNAs can be used as an efficient tool to engineer genomes to achieve large deletions or inversions, including fragments covering multiple genes and non-coding sequences.Abstract:
Customized TALENs and Cas9/gRNAs have been used for targeted mutagenesis in zebrafish to induce indels into protein-coding genes. However, indels are usually not sufficient to disrupt the function of non-coding genes, gene clusters or regulatory sequences, whereas large genomic deletions or inversions are more desirable for this purpose. By injecting two pairs of TALEN mRNAs or two gRNAs together with Cas9 mRNA targeting distal DNA sites of the same chromosome, we obtained predictable genomic deletions or inversions with sizes ranging from several hundred bases to nearly 1 Mb. We have successfully achieved this type of modifications for 11 chromosomal loci by TALENs and 2 by Cas9/gRNAs with different combinations of gRNA pairs, including clusters of miRNA and protein-coding genes. Seven of eight TALEN-targeted lines transmitted the deletions and one transmitted the inversion through germ line. Our findings indicate that both TALENs and Cas9/gRNAs can be used as an efficient tool to engineer genomes to achieve large deletions or inversions, including fragments covering multiple genes and non-coding sequences. To facilitate the analyses and application of existing ZFN, TALEN and CRISPR/Cas data, we have updated our EENdb database to provide a chromosomal view of all reported engineered endonucleases targeting human and zebrafish genomes.read more
Citations
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Journal ArticleDOI
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.
Conflict of interest statement. None declared.
TL;DR: It is found that women over 50 are more likely to have a family history of diabetes, especially if they are obese, than women under the age of 50.
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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
Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice
TL;DR: Adaptations of the type II CRISPR/Cas system leading to successful expression of the Cas9/sgRNA system in model plant and crop species bodes well for its near-term use as a facile and powerful means of plant genetic engineering for scientific and agricultural applications.
Journal ArticleDOI
A guide to genome engineering with programmable nucleases
Hyongbum Kim,Jin-Soo Kim +1 more
TL;DR: Known nuclease-specific features are essential for researchers to choose the most appropriate tool for a range of applications, including their composition, targetable sites, specificities and mutation signatures, among other characteristics.
References
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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.
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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.
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RNA-programmed genome editing in human cells
TL;DR: It is shown here that Cas9 assembles with hybrid guide RNAs in human cells and can induce the formation of double-strand DNA breaks at a site complementary to the guide RNA sequence in genomic DNA.
Journal ArticleDOI
Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease
TL;DR: It is shown that complexes of the Cas9 protein and artificial chimeric RNAs efficiently cleave two genomic sites and induce indels with frequencies of up to 33% in human cells.
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