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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235249/ https://www.ncbi.nlm.nih.gov/pubmed/32424114 http://dx.doi.org/10.1038/s41467-020-16117-8 |
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author | Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph M. Jakimo, Noah |
author_facet | Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph M. Jakimo, Noah |
author_sort | Chatterjee, Pranam |
collection | PubMed |
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[Formula: see text] -NAAN-3[Formula: see text] 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. |
format | Online Article Text |
id | pubmed-7235249 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-72352492020-05-20 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 M. Jakimo, Noah Nat Commun Article 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[Formula: see text] -NAAN-3[Formula: see text] 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. Nature Publishing Group UK 2020-05-18 /pmc/articles/PMC7235249/ /pubmed/32424114 http://dx.doi.org/10.1038/s41467-020-16117-8 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Chatterjee, Pranam Lee, Jooyoung Nip, Lisa Koseki, Sabrina R. T. Tysinger, Emma Sontheimer, Erik J. Jacobson, Joseph M. 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 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235249/ https://www.ncbi.nlm.nih.gov/pubmed/32424114 http://dx.doi.org/10.1038/s41467-020-16117-8 |
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