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Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries

The sequences of many genomes are available; therefore, relevant methods are needed for rapid and efficient identification of functional genes. The ability of tumour cells to resist apoptosis induced by anticancer agents may decide the success of failure of tumour elimination. Although the CD95-sign...

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Autores principales: Hassan, M, Matuschek, C, Gerber, PA, Peiper, M, Budach, W, Bölke, E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401000/
https://www.ncbi.nlm.nih.gov/pubmed/20564833
http://dx.doi.org/10.1186/2047-783X-15-4-162
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author Hassan, M
Matuschek, C
Gerber, PA
Peiper, M
Budach, W
Bölke, E
author_facet Hassan, M
Matuschek, C
Gerber, PA
Peiper, M
Budach, W
Bölke, E
author_sort Hassan, M
collection PubMed
description The sequences of many genomes are available; therefore, relevant methods are needed for rapid and efficient identification of functional genes. The ability of tumour cells to resist apoptosis induced by anticancer agents may decide the success of failure of tumour elimination. Although the CD95-signalling pathway is functional in tumour cells, the increased resistance of tumour cells to CD95-mediated apoptosis has been widely reported. In order to identify genes that might determine the response of tumour cells to CD95-mediated apoptosis, we modified the conventional technical knock out (TKO) strategy for isolation of genes that function in CD95-mediated apoptosis. Due to the fact that multiple different plasmids are usually introduced into the same cells, the effectiveness of the conventional TKO strategies is low. To overcome this obstacle, we replaced the conventional TKO strategy (based on stably expressed randomly fragmented cDNA libraries) with a multi-cycle selection procedure (based on transiently expressed randomly fragmented cDNA libraries with multi-cycle selection). Using this approach we could rapidly and significantly identify small numbers of antisense mRNA molecules, whose re-introduction into different tumour types confirmed their ability to block the pro-apoptotic function of their cognate genes. Thus, our modified TKO strategy provides a generally applicable procedure for the identification of functional genes with pro-apoptotic properties that may be clinically relevant to tumor therapy.
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spelling pubmed-34010002012-07-21 Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries Hassan, M Matuschek, C Gerber, PA Peiper, M Budach, W Bölke, E Eur J Med Res Review The sequences of many genomes are available; therefore, relevant methods are needed for rapid and efficient identification of functional genes. The ability of tumour cells to resist apoptosis induced by anticancer agents may decide the success of failure of tumour elimination. Although the CD95-signalling pathway is functional in tumour cells, the increased resistance of tumour cells to CD95-mediated apoptosis has been widely reported. In order to identify genes that might determine the response of tumour cells to CD95-mediated apoptosis, we modified the conventional technical knock out (TKO) strategy for isolation of genes that function in CD95-mediated apoptosis. Due to the fact that multiple different plasmids are usually introduced into the same cells, the effectiveness of the conventional TKO strategies is low. To overcome this obstacle, we replaced the conventional TKO strategy (based on stably expressed randomly fragmented cDNA libraries) with a multi-cycle selection procedure (based on transiently expressed randomly fragmented cDNA libraries with multi-cycle selection). Using this approach we could rapidly and significantly identify small numbers of antisense mRNA molecules, whose re-introduction into different tumour types confirmed their ability to block the pro-apoptotic function of their cognate genes. Thus, our modified TKO strategy provides a generally applicable procedure for the identification of functional genes with pro-apoptotic properties that may be clinically relevant to tumor therapy. BioMed Central 2010-04-08 /pmc/articles/PMC3401000/ /pubmed/20564833 http://dx.doi.org/10.1186/2047-783X-15-4-162 Text en Copyright ©2010 I. Holzapfel Publishers
spellingShingle Review
Hassan, M
Matuschek, C
Gerber, PA
Peiper, M
Budach, W
Bölke, E
Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title_full Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title_fullStr Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title_full_unstemmed Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title_short Identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented CDNA libraries
title_sort identification of candidate genes with pro-apoptotic properties by functional screening of randomly fragmented cdna libraries
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401000/
https://www.ncbi.nlm.nih.gov/pubmed/20564833
http://dx.doi.org/10.1186/2047-783X-15-4-162
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