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A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

Expression of the bacterial beta-galactosidase reporter gene (lacZ) in the vector used for the Knockout Mouse Project (KOMP) is driven by the endogenous promoter of the target gene. In tissues from KOMP mice, histochemical staining for LacZ enzyme activity can be used to determine gene expression pa...

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Autores principales: West, David B., Pasumarthi, Ravi K., Baridon, Brian, Djan, Esi, Trainor, Amanda, Griffey, Stephen M., Engelhard, Eric K., Rapp, Jared, Li, Bowen, de Jong, Pieter J., Lloyd, K.C. Kent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4381530/
https://www.ncbi.nlm.nih.gov/pubmed/25591789
http://dx.doi.org/10.1101/gr.184184.114
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author West, David B.
Pasumarthi, Ravi K.
Baridon, Brian
Djan, Esi
Trainor, Amanda
Griffey, Stephen M.
Engelhard, Eric K.
Rapp, Jared
Li, Bowen
de Jong, Pieter J.
Lloyd, K.C. Kent
author_facet West, David B.
Pasumarthi, Ravi K.
Baridon, Brian
Djan, Esi
Trainor, Amanda
Griffey, Stephen M.
Engelhard, Eric K.
Rapp, Jared
Li, Bowen
de Jong, Pieter J.
Lloyd, K.C. Kent
author_sort West, David B.
collection PubMed
description Expression of the bacterial beta-galactosidase reporter gene (lacZ) in the vector used for the Knockout Mouse Project (KOMP) is driven by the endogenous promoter of the target gene. In tissues from KOMP mice, histochemical staining for LacZ enzyme activity can be used to determine gene expression patterns. With this technique, we have produced a comprehensive resource of gene expression using both whole mount (WM) and frozen section (FS) LacZ staining in 313 unique KOMP mutant mouse lines. Of these, ∼80% of mutants showed specific staining in one or more tissues, while ∼20% showed no specific staining, ∼13% had staining in only one tissue, and ∼25% had staining in >6 tissues. The highest frequency of specific staining occurred in the brain (∼50%), male gonads (42%), and kidney (39%). The WM method was useful for rapidly identifying whole organ and some substructure staining, while the FS method often revealed substructure and cellular staining specificity. Both staining methods had >90% repeatability in biological replicates. Nonspecific LacZ staining occurs in some tissues due to the presence of bacteria or endogenous enzyme activity. However, this can be effectively distinguished from reporter gene activity by the combination of the WM and FS methods. After careful annotation, LacZ staining patterns in a high percentage of mutants revealed a unique structure-function not previously reported for many of these genes. The validation of methods for LacZ staining, annotation, and expression analysis reported here provides unique insights into the function of genes for which little is currently known.
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spelling pubmed-43815302015-10-01 A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines West, David B. Pasumarthi, Ravi K. Baridon, Brian Djan, Esi Trainor, Amanda Griffey, Stephen M. Engelhard, Eric K. Rapp, Jared Li, Bowen de Jong, Pieter J. Lloyd, K.C. Kent Genome Res Resource Expression of the bacterial beta-galactosidase reporter gene (lacZ) in the vector used for the Knockout Mouse Project (KOMP) is driven by the endogenous promoter of the target gene. In tissues from KOMP mice, histochemical staining for LacZ enzyme activity can be used to determine gene expression patterns. With this technique, we have produced a comprehensive resource of gene expression using both whole mount (WM) and frozen section (FS) LacZ staining in 313 unique KOMP mutant mouse lines. Of these, ∼80% of mutants showed specific staining in one or more tissues, while ∼20% showed no specific staining, ∼13% had staining in only one tissue, and ∼25% had staining in >6 tissues. The highest frequency of specific staining occurred in the brain (∼50%), male gonads (42%), and kidney (39%). The WM method was useful for rapidly identifying whole organ and some substructure staining, while the FS method often revealed substructure and cellular staining specificity. Both staining methods had >90% repeatability in biological replicates. Nonspecific LacZ staining occurs in some tissues due to the presence of bacteria or endogenous enzyme activity. However, this can be effectively distinguished from reporter gene activity by the combination of the WM and FS methods. After careful annotation, LacZ staining patterns in a high percentage of mutants revealed a unique structure-function not previously reported for many of these genes. The validation of methods for LacZ staining, annotation, and expression analysis reported here provides unique insights into the function of genes for which little is currently known. Cold Spring Harbor Laboratory Press 2015-04 /pmc/articles/PMC4381530/ /pubmed/25591789 http://dx.doi.org/10.1101/gr.184184.114 Text en © 2015 West et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Resource
West, David B.
Pasumarthi, Ravi K.
Baridon, Brian
Djan, Esi
Trainor, Amanda
Griffey, Stephen M.
Engelhard, Eric K.
Rapp, Jared
Li, Bowen
de Jong, Pieter J.
Lloyd, K.C. Kent
A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title_full A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title_fullStr A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title_full_unstemmed A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title_short A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines
title_sort lacz reporter gene expression atlas for 313 adult komp mutant mouse lines
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4381530/
https://www.ncbi.nlm.nih.gov/pubmed/25591789
http://dx.doi.org/10.1101/gr.184184.114
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