Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9
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 aureu...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Oxford University Press
2016-08-01
|
| Series: | G3: Genes, Genomes, Genetics |
| Subjects: | |
| Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.116.031914 |
| _version_ | 1819102038397550592 |
|---|---|
| author | Yan Feng Cheng Chen Yuxiang Han Zelin Chen Xiaochan Lu Fang Liang Song Li Wei Qin Shuo Lin |
| author_facet | Yan Feng Cheng Chen Yuxiang Han Zelin Chen Xiaochan Lu Fang Liang Song Li Wei Qin Shuo Lin |
| author_sort | Yan Feng |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T01:28:12Z |
| format | Article |
| id | doaj.art-9a927181f14a449bbe09c6db6d48fb3a |
| institution | Directory Open Access Journal |
| issn | 2160-1836 |
| language | English |
| last_indexed | 2024-12-22T01:28:12Z |
| publishDate | 2016-08-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | G3: Genes, Genomes, Genetics |
| spelling | doaj.art-9a927181f14a449bbe09c6db6d48fb3a2022-12-21T18:43:33ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362016-08-01682517252110.1534/g3.116.03191425Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9Yan FengCheng ChenYuxiang HanZelin ChenXiaochan LuFang LiangSong LiWei QinShuo LinThe 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.http://g3journal.org/lookup/doi/10.1534/g3.116.031914CRISPR/Cas9SaCas9KKH SaCas9 variantgene editingzebrafish |
| spellingShingle | Yan Feng Cheng Chen Yuxiang Han Zelin Chen Xiaochan Lu Fang Liang Song Li Wei Qin Shuo Lin Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 G3: Genes, Genomes, Genetics CRISPR/Cas9 SaCas9 KKH SaCas9 variant gene editing zebrafish |
| title | Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 |
| title_full | Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 |
| title_fullStr | Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 |
| title_full_unstemmed | Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 |
| title_short | Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 |
| title_sort | expanding crispr cas9 genome editing capacity in zebrafish using sacas9 |
| topic | CRISPR/Cas9 SaCas9 KKH SaCas9 variant gene editing zebrafish |
| url | http://g3journal.org/lookup/doi/10.1534/g3.116.031914 |
| work_keys_str_mv | AT yanfeng expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT chengchen expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT yuxianghan expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT zelinchen expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT xiaochanlu expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT fangliang expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT songli expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT weiqin expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 AT shuolin expandingcrisprcas9genomeeditingcapacityinzebrafishusingsacas9 |