Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells.
TDP-43 is found in cytoplasmic inclusions in 95% of amyotrophic lateral sclerosis (ALS) and 60% of frontotemporal lobar degeneration (FTLD). Approximately 4% of familial ALS is caused by mutations in TDP-43. The majority of these mutations are found in the glycine-rich domain, including the variant...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3961241?pdf=render |
| _version_ | 1828195451902361600 |
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| author | Agnes L Nishimura Carole Shum Emma L Scotter Amr Abdelgany Valentina Sardone Jamie Wright Youn-Bok Lee Han-Jou Chen Bilada Bilican Monica Carrasco Tom Maniatis Siddharthan Chandran Boris Rogelj Jean-Marc Gallo Christopher E Shaw |
| author_facet | Agnes L Nishimura Carole Shum Emma L Scotter Amr Abdelgany Valentina Sardone Jamie Wright Youn-Bok Lee Han-Jou Chen Bilada Bilican Monica Carrasco Tom Maniatis Siddharthan Chandran Boris Rogelj Jean-Marc Gallo Christopher E Shaw |
| author_sort | Agnes L Nishimura |
| collection | DOAJ |
| description | TDP-43 is found in cytoplasmic inclusions in 95% of amyotrophic lateral sclerosis (ALS) and 60% of frontotemporal lobar degeneration (FTLD). Approximately 4% of familial ALS is caused by mutations in TDP-43. The majority of these mutations are found in the glycine-rich domain, including the variant M337V, which is one of the most common mutations in TDP-43. In order to investigate the use of allele-specific RNA interference (RNAi) as a potential therapeutic tool, we designed and screened a set of siRNAs that specifically target TDP-43(M337V) mutation. Two siRNA specifically silenced the M337V mutation in HEK293T cells transfected with GFP-TDP-43(wt) or GFP-TDP-43(M337V) or TDP-43 C-terminal fragments counterparts. C-terminal TDP-43 transfected cells show an increase of cytosolic inclusions, which are decreased after allele-specific siRNA in M337V cells. We then investigated the effects of one of these allele-specific siRNAs in induced pluripotent stem cells (iPSCs) derived from an ALS patient carrying the M337V mutation. These lines showed a two-fold increase in cytosolic TDP-43 compared to the control. Following transfection with the allele-specific siRNA, cytosolic TDP-43 was reduced by 30% compared to cells transfected with a scrambled siRNA. We conclude that RNA interference can be used to selectively target the TDP-43(M337V) allele in mammalian and patient cells, thus demonstrating the potential for using RNA interference as a therapeutic tool for ALS. |
| first_indexed | 2024-04-12T09:42:47Z |
| format | Article |
| id | doaj.art-fa42f11aa6034af69f65a96cb5723752 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T09:42:47Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-fa42f11aa6034af69f65a96cb57237522022-12-22T03:38:02ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0193e9126910.1371/journal.pone.0091269Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells.Agnes L NishimuraCarole ShumEmma L ScotterAmr AbdelganyValentina SardoneJamie WrightYoun-Bok LeeHan-Jou ChenBilada BilicanMonica CarrascoTom ManiatisSiddharthan ChandranBoris RogeljJean-Marc GalloChristopher E ShawTDP-43 is found in cytoplasmic inclusions in 95% of amyotrophic lateral sclerosis (ALS) and 60% of frontotemporal lobar degeneration (FTLD). Approximately 4% of familial ALS is caused by mutations in TDP-43. The majority of these mutations are found in the glycine-rich domain, including the variant M337V, which is one of the most common mutations in TDP-43. In order to investigate the use of allele-specific RNA interference (RNAi) as a potential therapeutic tool, we designed and screened a set of siRNAs that specifically target TDP-43(M337V) mutation. Two siRNA specifically silenced the M337V mutation in HEK293T cells transfected with GFP-TDP-43(wt) or GFP-TDP-43(M337V) or TDP-43 C-terminal fragments counterparts. C-terminal TDP-43 transfected cells show an increase of cytosolic inclusions, which are decreased after allele-specific siRNA in M337V cells. We then investigated the effects of one of these allele-specific siRNAs in induced pluripotent stem cells (iPSCs) derived from an ALS patient carrying the M337V mutation. These lines showed a two-fold increase in cytosolic TDP-43 compared to the control. Following transfection with the allele-specific siRNA, cytosolic TDP-43 was reduced by 30% compared to cells transfected with a scrambled siRNA. We conclude that RNA interference can be used to selectively target the TDP-43(M337V) allele in mammalian and patient cells, thus demonstrating the potential for using RNA interference as a therapeutic tool for ALS.http://europepmc.org/articles/PMC3961241?pdf=render |
| spellingShingle | Agnes L Nishimura Carole Shum Emma L Scotter Amr Abdelgany Valentina Sardone Jamie Wright Youn-Bok Lee Han-Jou Chen Bilada Bilican Monica Carrasco Tom Maniatis Siddharthan Chandran Boris Rogelj Jean-Marc Gallo Christopher E Shaw Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. PLoS ONE |
| title | Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. |
| title_full | Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. |
| title_fullStr | Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. |
| title_full_unstemmed | Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. |
| title_short | Allele-specific knockdown of ALS-associated mutant TDP-43 in neural stem cells derived from induced pluripotent stem cells. |
| title_sort | allele specific knockdown of als associated mutant tdp 43 in neural stem cells derived from induced pluripotent stem cells |
| url | http://europepmc.org/articles/PMC3961241?pdf=render |
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