Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies
Granzyme A (GZMA) is a serine protease secreted by cytotoxic lymphocytes, with Gzma-/- mouse studies having informed our understanding of GZMA’s physiological function. We show herein that Gzma-/- mice have a mixed C57BL/6J and C57BL/6N genetic background and retain the full-length nicotinamide nucl...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-02-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/70207 |
| _version_ | 1828222199169810432 |
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| author | Daniel J Rawle Thuy T Le Troy Dumenil Cameron Bishop Kexin Yan Eri Nakayama Phillip I Bird Andreas Suhrbier |
| author_facet | Daniel J Rawle Thuy T Le Troy Dumenil Cameron Bishop Kexin Yan Eri Nakayama Phillip I Bird Andreas Suhrbier |
| author_sort | Daniel J Rawle |
| collection | DOAJ |
| description | Granzyme A (GZMA) is a serine protease secreted by cytotoxic lymphocytes, with Gzma-/- mouse studies having informed our understanding of GZMA’s physiological function. We show herein that Gzma-/- mice have a mixed C57BL/6J and C57BL/6N genetic background and retain the full-length nicotinamide nucleotide transhydrogenase (Nnt) gene, whereas Nnt is truncated in C57BL/6J mice. Chikungunya viral arthritis was substantially ameliorated in Gzma-/- mice; however, the presence of Nnt and the C57BL/6N background, rather than loss of GZMA expression, was responsible for this phenotype. A new CRISPR active site mutant C57BL/6J GzmaS211A mouse provided the first insights into GZMA’s bioactivity free of background issues, with circulating proteolytically active GZMA promoting immune-stimulating and pro-inflammatory signatures. Remarkably, k-mer mining of the Sequence Read Archive illustrated that ≈27% of Run Accessions and ≈38% of BioProjects listing C57BL/6J as the mouse strain had Nnt sequencing reads inconsistent with a C57BL/6J genetic background. Nnt and C57BL/6N background issues have clearly complicated our understanding of GZMA and may similarly have influenced studies across a broad range of fields. |
| first_indexed | 2024-04-12T16:51:45Z |
| format | Article |
| id | doaj.art-52beee401a264cc08b6f6799d457738e |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:51:45Z |
| publishDate | 2022-02-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-52beee401a264cc08b6f6799d457738e2022-12-22T03:24:23ZengeLife Sciences Publications LtdeLife2050-084X2022-02-011110.7554/eLife.70207Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studiesDaniel J Rawle0Thuy T Le1Troy Dumenil2https://orcid.org/0000-0002-3032-8360Cameron Bishop3https://orcid.org/0000-0002-5710-9942Kexin Yan4Eri Nakayama5Phillip I Bird6Andreas Suhrbier7https://orcid.org/0000-0001-8986-9025QIMR Berghofer Medical Research Institute, Brisbane, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, Australia; Department of Virology I, National Institute of Infectious Diseases, Tokyo, JapanDepartment of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, AustraliaQIMR Berghofer Medical Research Institute, Brisbane, Australia; Australian Infectious Disease Research Centre, GVN Center of Excellence, Brisbane, AustraliaGranzyme A (GZMA) is a serine protease secreted by cytotoxic lymphocytes, with Gzma-/- mouse studies having informed our understanding of GZMA’s physiological function. We show herein that Gzma-/- mice have a mixed C57BL/6J and C57BL/6N genetic background and retain the full-length nicotinamide nucleotide transhydrogenase (Nnt) gene, whereas Nnt is truncated in C57BL/6J mice. Chikungunya viral arthritis was substantially ameliorated in Gzma-/- mice; however, the presence of Nnt and the C57BL/6N background, rather than loss of GZMA expression, was responsible for this phenotype. A new CRISPR active site mutant C57BL/6J GzmaS211A mouse provided the first insights into GZMA’s bioactivity free of background issues, with circulating proteolytically active GZMA promoting immune-stimulating and pro-inflammatory signatures. Remarkably, k-mer mining of the Sequence Read Archive illustrated that ≈27% of Run Accessions and ≈38% of BioProjects listing C57BL/6J as the mouse strain had Nnt sequencing reads inconsistent with a C57BL/6J genetic background. Nnt and C57BL/6N background issues have clearly complicated our understanding of GZMA and may similarly have influenced studies across a broad range of fields.https://elifesciences.org/articles/70207granzyme Anicotinamide nucleotide transhydrogenasechikungunyainflammationC57BL/6JC57BL/6N |
| spellingShingle | Daniel J Rawle Thuy T Le Troy Dumenil Cameron Bishop Kexin Yan Eri Nakayama Phillip I Bird Andreas Suhrbier Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies eLife granzyme A nicotinamide nucleotide transhydrogenase chikungunya inflammation C57BL/6J C57BL/6N |
| title | Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies |
| title_full | Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies |
| title_fullStr | Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies |
| title_full_unstemmed | Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies |
| title_short | Widespread discrepancy in Nnt genotypes and genetic backgrounds complicates granzyme A and other knockout mouse studies |
| title_sort | widespread discrepancy in nnt genotypes and genetic backgrounds complicates granzyme a and other knockout mouse studies |
| topic | granzyme A nicotinamide nucleotide transhydrogenase chikungunya inflammation C57BL/6J C57BL/6N |
| url | https://elifesciences.org/articles/70207 |
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