Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9
| Autor: | Fang Liang, Yan Feng, Zelin Chen, Shuo Lin, Wei Qin, Yuxiang Han, Cheng Chen, Song Li, Xiaochan Lu |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Staphylococcus aureus Embryo Nonmammalian animal structures Bioinformatics analysis Genetically Modified QH426-470 Investigations Biology medicine.disease_cause Vertebrate Biology Genome Animals Genetically Modified 03 medical and health sciences 0302 clinical medicine Bacterial Proteins Genome editing Genetics medicine Animals KKH SaCas9 variant CRISPR CRISPR/Cas9 Molecular Biology Zebrafish Genetics (clinical) Gene Editing Nonmammalian Cas9 Human Genome fungi SaCas9 zebrafish biology.organism_classification Emerging Infectious Diseases 030104 developmental biology Embryo embryonic structures Streptococcus pyogenes CRISPR-Cas Systems 030217 neurology & neurosurgery Biotechnology |
| Zdroj: | G3 (Bethesda, Md.), vol 6, iss 8 G3: Genes|Genomes|Genetics G3: Genes, Genomes, Genetics, Vol 6, Iss 8, Pp 2517-2521 (2016) |
| ISSN: | 2160-1836 |
| DOI: | 10.1534/g3.116.031914 |
| Popis: | The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 5′-NGG-3′ protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 ortholog from Staphylococcus aureus (SaCas9), and its KKH variant, successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming previous findings, the SpCas9 variant, VQR, can also induce targeted mutations in zebrafish. Bioinformatics analysis of these new Cas targets suggests that the number of available target sites in the zebrafish genome can be greatly expanded. Collectively, the expanded target repertoire of Cas9 in zebrafish should further facilitate the utility of this organism for genetic studies of vertebrate biology. |
| Databáze: | OpenAIRE |
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