A Cas9 with PAM recognition for adenine dinucleotides
CRISPR-associated (Cas) DNA-endonucleases are remarkably effective tools for genome engineering, but have limited target ranges due to their protospacer adjacent motif (PAM) requirements. We demonstrate a critical expansion of the targetable sequence space for a type II-A CRISPR-associated enzyme th...
| Main Authors: | , , , , , , , |
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| Other Authors: | |
| Format: | Article |
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Springer Science and Business Media LLC
2020
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| Online Access: | https://hdl.handle.net/1721.1/125540 |
| _version_ | 1826208187050622976 |
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| author | Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph Jakimo, Noah |
| author2 | Massachusetts Institute of Technology. Center for Bits and Atoms |
| author_facet | Massachusetts Institute of Technology. Center for Bits and Atoms Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph Jakimo, Noah |
| author_sort | Chatterjee, Pranam |
| collection | MIT |
| description | CRISPR-associated (Cas) DNA-endonucleases are remarkably effective tools for genome engineering, but have limited target ranges due to their protospacer adjacent motif (PAM) requirements. We demonstrate a critical expansion of the targetable sequence space for a type II-A CRISPR-associated enzyme through identification of the natural 5′-NAAN-3′ PAM preference of Streptococcus macacae Cas9 (SmacCas9). To achieve efficient editing activity, we graft the PAM-interacting domain of SmacCas9 to its well-established ortholog from Streptococcus pyogenes (SpyCas9), and further engineer an increased efficiency variant (iSpyMac) for robust genome editing activity. We establish that our hybrids can target all adenine dinucleotide PAM sequences and possess robust and accurate editing capabilities in human cells. |
| first_indexed | 2024-09-23T14:01:53Z |
| format | Article |
| id | mit-1721.1/125540 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T14:01:53Z |
| publishDate | 2020 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1255402022-09-28T17:51:01Z A Cas9 with PAM recognition for adenine dinucleotides Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph Jakimo, Noah Massachusetts Institute of Technology. Center for Bits and Atoms Massachusetts Institute of Technology. Media Laboratory CRISPR-associated (Cas) DNA-endonucleases are remarkably effective tools for genome engineering, but have limited target ranges due to their protospacer adjacent motif (PAM) requirements. We demonstrate a critical expansion of the targetable sequence space for a type II-A CRISPR-associated enzyme through identification of the natural 5′-NAAN-3′ PAM preference of Streptococcus macacae Cas9 (SmacCas9). To achieve efficient editing activity, we graft the PAM-interacting domain of SmacCas9 to its well-established ortholog from Streptococcus pyogenes (SpyCas9), and further engineer an increased efficiency variant (iSpyMac) for robust genome editing activity. We establish that our hybrids can target all adenine dinucleotide PAM sequences and possess robust and accurate editing capabilities in human cells. 2020-05-28T13:47:51Z 2020-05-28T13:47:51Z 2020-05 2019-05 Article http://purl.org/eprint/type/JournalArticle 2041-1723 https://hdl.handle.net/1721.1/125540 Chatterjee, Pranam et al. "A Cas9 with PAM recognition for adenine dinucleotides." Nature Communications 11, 1 (May 2020): 2474 © 2020 Springer Nature http://dx.doi.org/10.1038/s41467-020-16117-8 Nature Communications Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Nature |
| spellingShingle | Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph Jakimo, Noah A Cas9 with PAM recognition for adenine dinucleotides |
| title | A Cas9 with PAM recognition for adenine dinucleotides |
| title_full | A Cas9 with PAM recognition for adenine dinucleotides |
| title_fullStr | A Cas9 with PAM recognition for adenine dinucleotides |
| title_full_unstemmed | A Cas9 with PAM recognition for adenine dinucleotides |
| title_short | A Cas9 with PAM recognition for adenine dinucleotides |
| title_sort | cas9 with pam recognition for adenine dinucleotides |
| url | https://hdl.handle.net/1721.1/125540 |
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