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Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system discovered using bacteria has been repurposed for genome editing in human cells. Transient expression of the editor proteins (e.g. Cas9 protein) is desirable to reduce the risk of mutagenesis from o...

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Autores principales: Lu, Baisong, Javidi-Parsijani, Parisa, Makani, Vishruti, Mehraein-Ghomi, Farideh, Sarhan, Walaa Mohamed, Sun, Dongjun, Yoo, Kyung Whan, Atala, Zachary P, Lyu, Pin, Atala, Anthony
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486560/
https://www.ncbi.nlm.nih.gov/pubmed/30759231
http://dx.doi.org/10.1093/nar/gkz093
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author Lu, Baisong
Javidi-Parsijani, Parisa
Makani, Vishruti
Mehraein-Ghomi, Farideh
Sarhan, Walaa Mohamed
Sun, Dongjun
Yoo, Kyung Whan
Atala, Zachary P
Lyu, Pin
Atala, Anthony
author_facet Lu, Baisong
Javidi-Parsijani, Parisa
Makani, Vishruti
Mehraein-Ghomi, Farideh
Sarhan, Walaa Mohamed
Sun, Dongjun
Yoo, Kyung Whan
Atala, Zachary P
Lyu, Pin
Atala, Anthony
author_sort Lu, Baisong
collection PubMed
description The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system discovered using bacteria has been repurposed for genome editing in human cells. Transient expression of the editor proteins (e.g. Cas9 protein) is desirable to reduce the risk of mutagenesis from off-target activity. Using the specific interaction between bacteriophage RNA-binding proteins and their RNA aptamers, we developed a system able to package up to 100 copies of Staphylococcus aureus Cas9 (SaCas9) mRNA in each lentivirus-like bionanoparticle (LVLP). The SaCas9 LVLPs mediated transient SaCas9 expression and achieved highly efficient genome editing in the presence of guide RNA. Lower off-target rates occurred in cells transduced with LVLPs containing SaCas9 mRNA, compared with cells transduced with adeno-associated virus or lentivirus expressing SaCas9. Our LVLP system may be useful for efficiently delivering Cas9 mRNA to cell lines and primary cells for in vitro and in vivo gene editing applications.
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spelling pubmed-64865602019-05-01 Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing Lu, Baisong Javidi-Parsijani, Parisa Makani, Vishruti Mehraein-Ghomi, Farideh Sarhan, Walaa Mohamed Sun, Dongjun Yoo, Kyung Whan Atala, Zachary P Lyu, Pin Atala, Anthony Nucleic Acids Res Methods Online The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system discovered using bacteria has been repurposed for genome editing in human cells. Transient expression of the editor proteins (e.g. Cas9 protein) is desirable to reduce the risk of mutagenesis from off-target activity. Using the specific interaction between bacteriophage RNA-binding proteins and their RNA aptamers, we developed a system able to package up to 100 copies of Staphylococcus aureus Cas9 (SaCas9) mRNA in each lentivirus-like bionanoparticle (LVLP). The SaCas9 LVLPs mediated transient SaCas9 expression and achieved highly efficient genome editing in the presence of guide RNA. Lower off-target rates occurred in cells transduced with LVLPs containing SaCas9 mRNA, compared with cells transduced with adeno-associated virus or lentivirus expressing SaCas9. Our LVLP system may be useful for efficiently delivering Cas9 mRNA to cell lines and primary cells for in vitro and in vivo gene editing applications. Oxford University Press 2019-05-07 2019-02-13 /pmc/articles/PMC6486560/ /pubmed/30759231 http://dx.doi.org/10.1093/nar/gkz093 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Lu, Baisong
Javidi-Parsijani, Parisa
Makani, Vishruti
Mehraein-Ghomi, Farideh
Sarhan, Walaa Mohamed
Sun, Dongjun
Yoo, Kyung Whan
Atala, Zachary P
Lyu, Pin
Atala, Anthony
Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title_full Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title_fullStr Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title_full_unstemmed Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title_short Delivering SaCas9 mRNA by lentivirus-like bionanoparticles for transient expression and efficient genome editing
title_sort delivering sacas9 mrna by lentivirus-like bionanoparticles for transient expression and efficient genome editing
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486560/
https://www.ncbi.nlm.nih.gov/pubmed/30759231
http://dx.doi.org/10.1093/nar/gkz093
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