About: Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells

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About: Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells
Creator	Acosta, Izani Blodorn, Eduardo Campos, Vinicius Corcini, Carine Daneluz, Larissa Dellagostin, Eduardo Domingues, William Fróes, Charles Komninou, Eliza Martins, Amanda Nunes, Leandro Rassier, Gabriela Varela, Antônio
Source	Medline; PMC
abstract	Sperm-mediated gene transfer (SMGT) has a potential use for zebrafish transgenesis. However, transfection into fish sperm cells still needs to be improved. The objective was to demonstrate the feasibility of tip type electroporation in zebrafish sperm, showing a protocol that provide high transfection efficiency, with minimal side-effects. Sperm was transfected with a Cy3-labelled DNA using tip type electroporation with voltages ranging from 500 to 1500 V. Sperm kinetics parameters were assessed using Computer Assisted Semen Analysis (CASA) and cell integrity, reactive oxygen species (ROS), mitochondrial functionality and transfection rate were evaluated by flow cytometry. The transfection rates were positively affected by tip type electroporation, reaching 64.9% ± 3.6 in the lowest voltage used (500 V) and 86.6% ± 1.9 in the highest (1500 V). The percentage of overall motile sperm in the electrotransfected samples was found to decrease with increasing field strength (P < 0.05). Increase in the sperm damaged plasma membrane was observed with increasing field strength (P < 0.05). ROS and sperm mitochondrial functionality did not present a negative response after the electroporation (P > 0.05). Overall results indicate that tip type electroporation enhances the internalization of exogenous DNA into zebrafish sperm cells with minimal harmful effects to sperm cells. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11033-020-05658-2) contains supplementary material, which is available to authorized users.
has issue date	2020-07-13(xsd:dateTime)
bibo:doi	10.1007/s11033-020-05658-2
bibo:pmid	32661869
has license	no-cc
sha1sum (hex)	e5c6f120f29fcf8bec0a8b4da7d01649046cd0f1
schema:url	https://doi.org/10.1007/s11033-020-05658-2
resource representing a document's title	Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells
has PubMed Central identifier	PMC7356131
has PubMed identifier	32661869
schema:publication	Mol Biol Rep
resource representing a document's body	covid:e5c6f120f29fcf8bec0a8b4da7d01649046cd0f1#body_text
is schema:about of	named entity 'flow cytometry' named entity 'provide' named entity 'damaged' named entity 'protocol' named entity 'high' named entity 'reactive oxygen species (ROS)' named entity 'internalization' named entity 'sperm cells' named entity 'sperm cells' named entity 'transfection' named entity 'DID' named entity 'EXOGENOUS' named entity 'SEMEN ANALYSIS' named entity 'TRANSFECTION EFFICIENCY' named entity 'TYPE' named entity 'FUNCTIONALITY' named entity 'MITOCHONDRIAL' named entity 'EVALUATED' named entity 'FIELD STRENGTH' named entity 'DNA' named entity 'HARMFUL' named entity 'minimal' named entity 'objective' named entity 'assessed' named entity 'Analysis' named entity 'cell' named entity 'field strength' named entity 'DNA' named entity 'reaching' named entity 'transfected' named entity 'transfection' named entity 'Computer' named entity 'ROS' named entity 'reactive oxygen species' named entity 'transgenesis' named entity 'motile' named entity 'transfection' named entity 'sperm' named entity 'harmful effects' named entity 'Cy3' named entity 'sperm' named entity 'transfection' named entity 'plasma membrane' named entity 'transfection' named entity 'Sperm' named entity 'voltage' named entity 'sperm cells' named entity 'sperm' named entity 'sperm cells' named entity 'sperm cells' named entity 'cell motility' named entity 'sperm parameters' named entity 'cell viability' named entity 'electroporation' named entity 'transfected' named entity 'exogenous DNA' named entity 'sperm' named entity 'electroporation' named entity 'Shapiro-Wilk test' named entity 'STR' named entity 'fertilization' named entity 'Sperm' named entity 'sperm' named entity 'Mitochondrial function' named entity 'mitochondria' named entity 'transfection' named entity 'DNase' named entity 'transfection' named entity 'electroporation'

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