CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation

Abstract The imprinted H19 long non-coding RNA, a knowing oncofetal gene, presents a controversial role during the carcinogenesis process since its tumor suppressor or oncogenic activity is not completely elucidated. Since H19 lncRNA is involved in many biological pathways related to tumorigenesis,...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Renan da Silva Santos, Daniel Pascoalino Pinheiro, Louhanna Pinheiro Rodrigues Teixeira, Sarah Leyenne Alves Sales, Maria Claudia dos Santos Luciano, Mayara Magna de Lima Melo, Ronald Feitosa Pinheiro, Kaio César Simiano Tavares, Gilvan Pessoa Furtado, Claudia Pessoa, Cristiana Libardi Miranda Furtado
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fa96d963cb0e487eafea13981072274b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:fa96d963cb0e487eafea13981072274b
record_format	dspace
spelling	oai:doaj.org-article:fa96d963cb0e487eafea13981072274b2021-12-02T15:16:05ZCRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation10.1038/s41598-021-97058-02045-2322https://doaj.org/article/fa96d963cb0e487eafea13981072274b2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97058-0https://doaj.org/toc/2045-2322Abstract The imprinted H19 long non-coding RNA, a knowing oncofetal gene, presents a controversial role during the carcinogenesis process since its tumor suppressor or oncogenic activity is not completely elucidated. Since H19 lncRNA is involved in many biological pathways related to tumorigenesis, we sought to develop a non-cancer lineage with CRISPR-Cas9-mediated H19 knockdown (H19-) and observe the changes in a cellular context. To edit the promoter region of H19, two RNA guides were designed, and the murine C2C12 myoblast cells were transfected. H19 deletion was determined by DNA sequencing and gene expression by qPCR. We observed a small deletion (~ 60 bp) in the promoter region that presented four predicted transcription binding sites. The deletion reduced H19 expression (30%) and resulted in increased proliferative activity, altered morphological patterns including cell size and intracellular granularity, without changes in viability. The increased proliferation rate in the H19- cell seems to facilitate chromosomal abnormalities. The H19- myoblast presented characteristics similar to cancer cells, therefore the H19 lncRNA may be an important gene during the initiation of the tumorigenic process. Due to CRISPR/Cas9 permanent edition, the C2C12 H19- knockdown cells allows functional studies of H19 roles in tumorigenesis, prognosis, metastases, as well as drug resistance and targeted therapy.Renan da Silva SantosDaniel Pascoalino PinheiroLouhanna Pinheiro Rodrigues TeixeiraSarah Leyenne Alves SalesMaria Claudia dos Santos LucianoMayara Magna de Lima MeloRonald Feitosa PinheiroKaio César Simiano TavaresGilvan Pessoa FurtadoClaudia PessoaCristiana Libardi Miranda FurtadoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Renan da Silva Santos Daniel Pascoalino Pinheiro Louhanna Pinheiro Rodrigues Teixeira Sarah Leyenne Alves Sales Maria Claudia dos Santos Luciano Mayara Magna de Lima Melo Ronald Feitosa Pinheiro Kaio César Simiano Tavares Gilvan Pessoa Furtado Claudia Pessoa Cristiana Libardi Miranda Furtado CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
description	Abstract The imprinted H19 long non-coding RNA, a knowing oncofetal gene, presents a controversial role during the carcinogenesis process since its tumor suppressor or oncogenic activity is not completely elucidated. Since H19 lncRNA is involved in many biological pathways related to tumorigenesis, we sought to develop a non-cancer lineage with CRISPR-Cas9-mediated H19 knockdown (H19-) and observe the changes in a cellular context. To edit the promoter region of H19, two RNA guides were designed, and the murine C2C12 myoblast cells were transfected. H19 deletion was determined by DNA sequencing and gene expression by qPCR. We observed a small deletion (~ 60 bp) in the promoter region that presented four predicted transcription binding sites. The deletion reduced H19 expression (30%) and resulted in increased proliferative activity, altered morphological patterns including cell size and intracellular granularity, without changes in viability. The increased proliferation rate in the H19- cell seems to facilitate chromosomal abnormalities. The H19- myoblast presented characteristics similar to cancer cells, therefore the H19 lncRNA may be an important gene during the initiation of the tumorigenic process. Due to CRISPR/Cas9 permanent edition, the C2C12 H19- knockdown cells allows functional studies of H19 roles in tumorigenesis, prognosis, metastases, as well as drug resistance and targeted therapy.
format	article
author	Renan da Silva Santos Daniel Pascoalino Pinheiro Louhanna Pinheiro Rodrigues Teixeira Sarah Leyenne Alves Sales Maria Claudia dos Santos Luciano Mayara Magna de Lima Melo Ronald Feitosa Pinheiro Kaio César Simiano Tavares Gilvan Pessoa Furtado Claudia Pessoa Cristiana Libardi Miranda Furtado
author_facet	Renan da Silva Santos Daniel Pascoalino Pinheiro Louhanna Pinheiro Rodrigues Teixeira Sarah Leyenne Alves Sales Maria Claudia dos Santos Luciano Mayara Magna de Lima Melo Ronald Feitosa Pinheiro Kaio César Simiano Tavares Gilvan Pessoa Furtado Claudia Pessoa Cristiana Libardi Miranda Furtado
author_sort	Renan da Silva Santos
title	CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
title_short	CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
title_full	CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
title_fullStr	CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
title_full_unstemmed	CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation
title_sort	crispr/cas9 small promoter deletion in h19 lncrna is associated with altered cell morphology and proliferation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/fa96d963cb0e487eafea13981072274b
work_keys_str_mv	AT renandasilvasantos crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT danielpascoalinopinheiro crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT louhannapinheirorodriguesteixeira crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT sarahleyennealvessales crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT mariaclaudiadossantosluciano crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT mayaramagnadelimamelo crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT ronaldfeitosapinheiro crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT kaiocesarsimianotavares crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT gilvanpessoafurtado crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT claudiapessoa crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation AT cristianalibardimirandafurtado crisprcas9smallpromoterdeletioninh19lncrnaisassociatedwithalteredcellmorphologyandproliferation
_version_	1718387547517747200

CRISPR/Cas9 small promoter deletion in H19 lncRNA is associated with altered cell morphology and proliferation

Ejemplares similares