Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation.
Targeted mutations in mouse disrupt local chromatin structure and may lead to unanticipated local effects. We evaluated targeted gene promoter silencing in a group of six mutants carrying the tm1a Knockout Mouse Project allele containing both a LacZ reporter gene driven by the native promoter and a...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4537176?pdf=render |
| _version_ | 1818318748202827776 |
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| author | Julia V Kirov Michael Adkisson A J Nava Andreana Cipollone Brandon Willis Eric K Engelhard K C Kent Lloyd Pieter de Jong David B West |
| author_facet | Julia V Kirov Michael Adkisson A J Nava Andreana Cipollone Brandon Willis Eric K Engelhard K C Kent Lloyd Pieter de Jong David B West |
| author_sort | Julia V Kirov |
| collection | DOAJ |
| description | Targeted mutations in mouse disrupt local chromatin structure and may lead to unanticipated local effects. We evaluated targeted gene promoter silencing in a group of six mutants carrying the tm1a Knockout Mouse Project allele containing both a LacZ reporter gene driven by the native promoter and a neo selection cassette. Messenger RNA levels of the reporter gene and targeted gene were assessed by qRT-PCR, and methylation of the promoter CpG islands and LacZ coding sequence were evaluated by sequencing of bisulfite-treated DNA. Mutants were stratified by LacZ staining into presumed Silenced and Expressed reporter genes. Silenced mutants had reduced relative quantities LacZ mRNA and greater CpG Island methylation compared with the Expressed mutant group. Within the silenced group, LacZ coding sequence methylation was significantly and positively correlated with CpG Island methylation, while promoter CpG methylation was only weakly correlated with LacZ gene mRNA. The results support the conclusion that there is promoter silencing in a subset of mutants carrying the tm1a allele. The features of targeted genes which promote local silencing when targeted remain unknown. |
| first_indexed | 2024-12-13T09:58:09Z |
| format | Article |
| id | doaj.art-bb67362f1ea14d198d4793e888e3eb46 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T09:58:09Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-bb67362f1ea14d198d4793e888e3eb462022-12-21T23:51:44ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01108e013415510.1371/journal.pone.0134155Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation.Julia V KirovMichael AdkissonA J NavaAndreana CipolloneBrandon WillisEric K EngelhardK C Kent LloydPieter de JongDavid B WestTargeted mutations in mouse disrupt local chromatin structure and may lead to unanticipated local effects. We evaluated targeted gene promoter silencing in a group of six mutants carrying the tm1a Knockout Mouse Project allele containing both a LacZ reporter gene driven by the native promoter and a neo selection cassette. Messenger RNA levels of the reporter gene and targeted gene were assessed by qRT-PCR, and methylation of the promoter CpG islands and LacZ coding sequence were evaluated by sequencing of bisulfite-treated DNA. Mutants were stratified by LacZ staining into presumed Silenced and Expressed reporter genes. Silenced mutants had reduced relative quantities LacZ mRNA and greater CpG Island methylation compared with the Expressed mutant group. Within the silenced group, LacZ coding sequence methylation was significantly and positively correlated with CpG Island methylation, while promoter CpG methylation was only weakly correlated with LacZ gene mRNA. The results support the conclusion that there is promoter silencing in a subset of mutants carrying the tm1a allele. The features of targeted genes which promote local silencing when targeted remain unknown.http://europepmc.org/articles/PMC4537176?pdf=render |
| spellingShingle | Julia V Kirov Michael Adkisson A J Nava Andreana Cipollone Brandon Willis Eric K Engelhard K C Kent Lloyd Pieter de Jong David B West Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. PLoS ONE |
| title | Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. |
| title_full | Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. |
| title_fullStr | Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. |
| title_full_unstemmed | Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. |
| title_short | Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation. |
| title_sort | reporter gene silencing in targeted mouse mutants is associated with promoter cpg island methylation |
| url | http://europepmc.org/articles/PMC4537176?pdf=render |
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