Highly efficient serum-free manipulation of miRNA in human NK cells without loss of viability or phenotypic alterations is accomplished with TransIT-TKO

Breanna K.V. Hargreaves; Sarah E. Roberts; Beata Derfalvi; Jeanette E. Boudreau

doi:10.1371/journal.pone.0231664

Highly efficient serum-free manipulation of miRNA in human NK cells without loss of viability or phenotypic alterations is accomplished with TransIT-TKO

Breanna K.V. Hargreaves, Sarah E. Roberts, Beata Derfalvi, Jeanette E. Boudreau

Medicine

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9 Citas (Scopus)

Resumen

Natural killer (NK) cells are innate lymphocytes with functions that include target cell killing, inflammation and regulation. NK cells integrate incoming activating and inhibitory signals through an array of germline-encoded receptors to gauge the health of neighbouring cells. The reactive potential of NK cells is influenced by microRNA (miRNA), small non-coding sequences that interfere with mRNA expression. miRNAs are highly conserved between species, and a single miRNA can have hundreds to thousands of targets and influence entire cellular programs. Two miRNA species, miR-155-5p and miR-146a-5p are known to be important in controlling NK cell function, but research to best understand the impacts of miRNA species within NK cells has been bottlenecked by a lack of techniques for altering miRNA concentrations efficiently and without off-target effects. Here, we describe a non-viral and straightforward approach for increasing or decreasing expression of miRNA in primary human NK cells. We achieve >90% transfection efficiency without off-target impacts on NK cell viability, education, phenotype or function. This opens the opportunity to study and manipulate NK cell miRNA profiles and their impacts on NK cellular programs which may influence outcomes of cancer, inflammation and autoimmunity.

Idioma original	English
Número de artículo	e0231664
Publicación	PLoS One
Volumen	15
N.º	4
DOI	https://doi.org/10.1371/journal.pone.0231664
Estado	Published - abr. 2020

Nota bibliográfica

Funding Information:
This work was supported by studentships to BKVH from the Nova Scotia Graduate Scholarship and the Nova Scotia Health Research (NSHRF) foundation, and grants to BD (Canada Foundation for Innovation, John R. Evans Leaders Fund #34283, NSHRF Establishment Grant #1045, IWK Establishment Grant 2017, Dalhousie University Dept. of Pediatrics Endowment Fund, 2018), the Natural Sciences and Engineering Research Council #06489 of Canada to JEB, and Canadian Institutes of Health Research (CIHR), REACH Program Team Grant: Health Challenges in Chronic Inflammation Initiative (CIHR 135230). The sponsors did not play a role in any aspect of the study. We sincerely thank the healthy blood donors for their contribution to this project and gratefully acknowledge the Dalhousie University Faculty of Medicine Flow Cytometry CORE facility, Derek Rowter, Renee Raudonis and Fang Liu for technical assistance with this project.

Publisher Copyright:
© 2020 Hargreaves et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Journal Article
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title = "Highly efficient serum-free manipulation of miRNA in human NK cells without loss of viability or phenotypic alterations is accomplished with TransIT-TKO",

abstract = "Natural killer (NK) cells are innate lymphocytes with functions that include target cell killing, inflammation and regulation. NK cells integrate incoming activating and inhibitory signals through an array of germline-encoded receptors to gauge the health of neighbouring cells. The reactive potential of NK cells is influenced by microRNA (miRNA), small non-coding sequences that interfere with mRNA expression. miRNAs are highly conserved between species, and a single miRNA can have hundreds to thousands of targets and influence entire cellular programs. Two miRNA species, miR-155-5p and miR-146a-5p are known to be important in controlling NK cell function, but research to best understand the impacts of miRNA species within NK cells has been bottlenecked by a lack of techniques for altering miRNA concentrations efficiently and without off-target effects. Here, we describe a non-viral and straightforward approach for increasing or decreasing expression of miRNA in primary human NK cells. We achieve >90% transfection efficiency without off-target impacts on NK cell viability, education, phenotype or function. This opens the opportunity to study and manipulate NK cell miRNA profiles and their impacts on NK cellular programs which may influence outcomes of cancer, inflammation and autoimmunity.",

author = "Hargreaves, {Breanna K.V.} and Roberts, {Sarah E.} and Beata Derfalvi and Boudreau, {Jeanette E.}",

note = "Funding Information: This work was supported by studentships to BKVH from the Nova Scotia Graduate Scholarship and the Nova Scotia Health Research (NSHRF) foundation, and grants to BD (Canada Foundation for Innovation, John R. Evans Leaders Fund #34283, NSHRF Establishment Grant #1045, IWK Establishment Grant 2017, Dalhousie University Dept. of Pediatrics Endowment Fund, 2018), the Natural Sciences and Engineering Research Council #06489 of Canada to JEB, and Canadian Institutes of Health Research (CIHR), REACH Program Team Grant: Health Challenges in Chronic Inflammation Initiative (CIHR 135230). The sponsors did not play a role in any aspect of the study. We sincerely thank the healthy blood donors for their contribution to this project and gratefully acknowledge the Dalhousie University Faculty of Medicine Flow Cytometry CORE facility, Derek Rowter, Renee Raudonis and Fang Liu for technical assistance with this project. Publisher Copyright: {\textcopyright} 2020 Hargreaves et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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AU - Boudreau, Jeanette E.

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Highly efficient serum-free manipulation of miRNA in human NK cells without loss of viability or phenotypic alterations is accomplished with TransIT-TKO

Resumen

Nota bibliográfica

ASJC Scopus Subject Areas

PubMed: MeSH publication types

ODS de las Naciones Unidas

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