Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection

Exploring the essential role of Importin 11 (IPO11) in the nuclear translocation of its potential cargo proteins requires an efficient means of IPO11 deletion and re-expression. Here, we present a protocol for the generation of IPO11 deletion using CRISPR-Cas9 and re-expression using plasmid transfe...

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Detalles Bibliográficos
Autores principales: Zhang, Yarui, Ou, Guomin, Zhou, Zhou, Wang, Jingdong, Yang, Hongying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10206203/
https://www.ncbi.nlm.nih.gov/pubmed/37195868
http://dx.doi.org/10.1016/j.xpro.2023.102317
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author Zhang, Yarui
Ou, Guomin
Zhou, Zhou
Wang, Jingdong
Yang, Hongying
author_facet Zhang, Yarui
Ou, Guomin
Zhou, Zhou
Wang, Jingdong
Yang, Hongying
author_sort Zhang, Yarui
collection PubMed
description Exploring the essential role of Importin 11 (IPO11) in the nuclear translocation of its potential cargo proteins requires an efficient means of IPO11 deletion and re-expression. Here, we present a protocol for the generation of IPO11 deletion using CRISPR-Cas9 and re-expression using plasmid transfection in H460 non-small cell lung cancer cells. We describe steps for lentiviral transduction of H460 cells, single clone selection, and expansion and validation of cell colonies. We then detail plasmid transfection and validation of transfection efficiency. For complete details on the use and execution of this protocol, please refer to Zhang et al.(1)
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spelling pubmed-102062032023-05-25 Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection Zhang, Yarui Ou, Guomin Zhou, Zhou Wang, Jingdong Yang, Hongying STAR Protoc Protocol Exploring the essential role of Importin 11 (IPO11) in the nuclear translocation of its potential cargo proteins requires an efficient means of IPO11 deletion and re-expression. Here, we present a protocol for the generation of IPO11 deletion using CRISPR-Cas9 and re-expression using plasmid transfection in H460 non-small cell lung cancer cells. We describe steps for lentiviral transduction of H460 cells, single clone selection, and expansion and validation of cell colonies. We then detail plasmid transfection and validation of transfection efficiency. For complete details on the use and execution of this protocol, please refer to Zhang et al.(1) Elsevier 2023-05-16 /pmc/articles/PMC10206203/ /pubmed/37195868 http://dx.doi.org/10.1016/j.xpro.2023.102317 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Zhang, Yarui
Ou, Guomin
Zhou, Zhou
Wang, Jingdong
Yang, Hongying
Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title_full Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title_fullStr Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title_full_unstemmed Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title_short Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection
title_sort protocol for ipo11 deletion and re-expression in h460 lung cancer cells using crispr-cas9 and plasmid transfection
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10206203/
https://www.ncbi.nlm.nih.gov/pubmed/37195868
http://dx.doi.org/10.1016/j.xpro.2023.102317
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