Publication:

Cellular transfection of small interfering ribonucleic acids. Electroporation as an alternative to liposome-based delivery transfection methods

cris.legacyId

155051

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PTCB

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0000-0003-0139-223X

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EPFL

cris.virtual.parent-organization

ETU

cris.virtual.parent-organization

EPFL

cris.virtual.sciperId

103994

cris.virtual.unitManager

Turcatti, Gerardo

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7573884a-489f-4154-aeed-d8fc5b340a7a

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7573884a-489f-4154-aeed-d8fc5b340a7a

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7573884a-489f-4154-aeed-d8fc5b340a7a

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36cdc379-b0ee-43e7-b0f2-261f07274461

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d026b261-5306-4998-b7f2-7057fdbec621

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d026b261-5306-4998-b7f2-7057fdbec621

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e95b5577-48a6-4034-9fe5-cb5a78e1bb28

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e95b5577-48a6-4034-9fe5-cb5a78e1bb28

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e95b5577-48a6-4034-9fe5-cb5a78e1bb28

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7573884a-489f-4154-aeed-d8fc5b340a7a

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7573884a-489f-4154-aeed-d8fc5b340a7a

cris.virtualsource.unitId

36cdc379-b0ee-43e7-b0f2-261f07274461

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36cdc379-b0ee-43e7-b0f2-261f07274461

cris.virtualsource.unitManager

7510ba3a-086b-4be9-85f8-e43d2ca5d91f

datacite.rights

metadata-only

dc.contributor.advisor

Turcatti, Gerardo

dc.contributor.author

Jacot, Guillaume

dc.date.accessioned

2010-11-18T15:26:43

dc.date.available

2010-11-18T15:26:43

dc.date.created

2010-11-18

dc.date.issued

2010

dc.date.modified

2025-01-23T10:40:32.550265Z

dc.description.abstract

RNA interference has become an increasingly important tool for all aspects of molecular biology. Nevertheless, there is a technological challenge for performing gene knock-down with chemical transfection agents, as many relevant cell types are refractory to efficient transport of short interfering RNA (siRNA) into cytosol using these classical carrier-based methods. The aim of this project was to evaluate the potential of electroporation as a generic transfection method and to compare it with a standard and well-established chemical reversetransfection using cationic lipids as carriers for short-interfering RNAs. We have used an automated cell electroporation instrument from a swedish company, Cellectricon. The Cellaxess HT instrument is able to perform electroporation in 384 wells plates. We compared cell viability, transfection efficiency and extend of gene silencing caused by electroporation, with reverse-transfection using Lipofectamine RNAiMAX. Gene knock down efficiency has been measured at the protein function level by targeting ubiquitously expressed genes that are essential to cell survival. Cell viability and transfection efficiency are, then, opposed with this strategy. Results clearly showed a net improvement of "difficult-to-transfect" cell transfection but this improvement comes at the price of consuming up to ten fold more siRNAs. For "normal" cell line, lipofection remains the preferred methodology as it is more efficient and less expensive. Furthermore, the Cellaxess HT instrument has some serious drawbacks that pervert easy and reproducible result acquisition.

dc.description.notes

Biomolecular Screening Facility, EPFL

dc.description.sponsorship

SSV

dc.identifier.uri

https://infoscience.epfl.ch/handle/20.500.14299/57785

dc.subject

Bioingénierie et Biotechnologie

dc.subject

Bioengineering and Biotechnology

dc.subject

FSV/SSV

dc.title

Cellular transfection of small interfering ribonucleic acids. Electroporation as an alternative to liposome-based delivery transfection methods

dc.type

student work::master thesis

dspace.entity.type

Publication

dspace.legacy.oai-identifier

oai:infoscience.tind.io:155051

epfl.legacy.itemtype

Student Projects

epfl.legacy.submissionform

STUDENT

epfl.oai.currentset

OpenAIREv4

epfl.oai.currentset

SV

epfl.thesis.degreelevel

Master's Thesis

epfl.thesis.faculty

SV

epfl.thesis.section

SV-S

epfl.writtenAt

EPFL

oairecerif.advisor.affiliation

EPFL

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