CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.

With the ever-increasing developing rate of gene and cellular therapy applications and growing accessibility due to products receiving regulatory approval, the need for effective and reliable safety mechanisms to prevent or eliminate potentially fatal side effects is of the utmost importance. In thi...

ver descrição completa

Detalhes bibliográficos
Autores: Amberger, Maximilian, Grueso Hierro, Mª Esther, Ivics, Zoltán
Tipo de documento: artigo
Data de publicação:2023
País:España
Recursos:Universidad Francisco de Vitoria
Repositório:DDFV. Repositorio Institucional de la Universidad Francisco de Vitoria
Idioma:inglês
OAI Identifier:oai:ddfv.ufv.es:10641/5625
Acesso em linha:https://hdl.handle.net/10641/5625
Access Level:Acceso aberto
Palavra-chave:Suicide switch
Suicide gene
CRISPR/Cas9
Alu retrotransposon
Sleeping Beauty transposon
Gene therapy
id ES_6fa1f9c6abef80b99dcdf90d7e1e7eab
oai_identifier_str oai:ddfv.ufv.es:10641/5625
network_acronym_str ES
network_name_str España
spelling CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch. Amberger, Maximilian Grueso Hierro, Mª Esther Ivics, Zoltán Suicide switch Suicide gene CRISPR/Cas9 Alu retrotransposon Sleeping Beauty transposon Gene therapy With the ever-increasing developing rate of gene and cellular therapy applications and growing accessibility due to products receiving regulatory approval, the need for effective and reliable safety mechanisms to prevent or eliminate potentially fatal side effects is of the utmost importance. In this study, we present the CRISPR-induced suicide switch (CRISISS) as a tool to eliminate genetically modified cells in an inducible and highly efficient manner by targeting Cas9 to highly repetitive Alu retrotransposons in the human genome, causing irreparable genomic fragmentation by the Cas9 nuclease and resulting cell death. The suicide switch components, including expression cassettes for a transcriptionally and post-translationally inducible Cas9 and an Alu-specific single-guide RNA, were integrated into the genome of target cells via Sleeping-Beauty-mediated transposition. The resulting transgenic cells did not show signs of any impact on overall fitness when uninduced, as unintended background expression, background DNA damage response and background cell killing were not observed. When induced, however, a strong expression of Cas9, a strong DNA damage response and a rapid halt of cell proliferation coupled with near complete cell death within four days post-induction were seen. With this proof-of-concept study, we present a novel and promising approach for a robust suicide switch with potential utility for gene and cell therapy in the future. MDPI https://hdl.handle.net/10641/5625
title CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
spellingShingle CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
Amberger, Maximilian
Suicide switch
Suicide gene
CRISPR/Cas9
Alu retrotransposon
Sleeping Beauty transposon
Gene therapy
title_short CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
title_full CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
title_fullStr CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
title_full_unstemmed CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
title_sort CRISISS: a novel, transcriptionally and post-translationally inducible CRISPR/Cas9-based cellular suicide switch.
author Amberger, Maximilian
author_facet Amberger, Maximilian
Grueso Hierro, Mª Esther
Ivics, Zoltán
author_role author
author2 Grueso Hierro, Mª Esther
Ivics, Zoltán
author2_role author
author
topic Suicide switch
Suicide gene
CRISPR/Cas9
Alu retrotransposon
Sleeping Beauty transposon
Gene therapy
topic_facet Suicide switch
Suicide gene
CRISPR/Cas9
Alu retrotransposon
Sleeping Beauty transposon
Gene therapy
description With the ever-increasing developing rate of gene and cellular therapy applications and growing accessibility due to products receiving regulatory approval, the need for effective and reliable safety mechanisms to prevent or eliminate potentially fatal side effects is of the utmost importance. In this study, we present the CRISPR-induced suicide switch (CRISISS) as a tool to eliminate genetically modified cells in an inducible and highly efficient manner by targeting Cas9 to highly repetitive Alu retrotransposons in the human genome, causing irreparable genomic fragmentation by the Cas9 nuclease and resulting cell death. The suicide switch components, including expression cassettes for a transcriptionally and post-translationally inducible Cas9 and an Alu-specific single-guide RNA, were integrated into the genome of target cells via Sleeping-Beauty-mediated transposition. The resulting transgenic cells did not show signs of any impact on overall fitness when uninduced, as unintended background expression, background DNA damage response and background cell killing were not observed. When induced, however, a strong expression of Cas9, a strong DNA damage response and a rapid halt of cell proliferation coupled with near complete cell death within four days post-induction were seen. With this proof-of-concept study, we present a novel and promising approach for a robust suicide switch with potential utility for gene and cell therapy in the future.
publishDate 2023
format article
url https://hdl.handle.net/10641/5625
language eng
eu_rights_str_mv openAccess
publisher MDPI
institution Universidad Francisco de Vitoria
collection DDFV. Repositorio Institucional de la Universidad Francisco de Vitoria
reponame_str DDFV. Repositorio Institucional de la Universidad Francisco de Vitoria
instname_str Universidad Francisco de Vitoria
_version_ 1878436172963774464
publishDateSort 2023
author_browse Amberger, Maximilian
Grueso Hierro, Mª Esther
Ivics, Zoltán
publisherStr MDPI
score 6,924472

Registros relacionados