Open AccessKnockout of ISCA 1 causes early embryonic death in rats
Author(s) -
Yang Xinlan,
Lu Dan,
Zhang Xu,
Chen Wei,
Gao Shan,
Dong Wei,
Ma Yuanwu,
Zhang Lianfeng
Publication year - 2019
Publication title -
animal models and experimental medicine
Language(s) - English
Resource type - Journals
ISSN - 2576-2095
DOI - 10.1002/ame2.12059
Subject(s) - knockout mouse , chemistry , microbiology and biotechnology , gene , biology , biochemistry
Abstract Background Iron‐sulfur cluster assembly 1 ( ISCA 1) is an iron‐sulfur (Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing proteins. ISCA 1 is also the newly identified causal gene for multiple mitochondrial dysfunctions syndrome ( MMDS ). However, our knowledge about the physiological function of ISCA 1 in vivo is currently limited. In this study, we generated an ISCA 1 knockout rat line and analyzed the embryo development. Methods ISCA 1 knockout rats were generated by replacing the exon1 of ISCA 1 gene with the mC herry‐Cre fusion gene using CRISPR ‐Cas9 technology. The ISCA 1 expression pattern was analyzed by fluorescence imaging using ISCA 1 promotor driven Cre and mC herry expression. The embryonic morphology was examinated by microscope and mitochondrial proteins were tested by Western blot. Results An ISCA 1 knockout rat line was obtained, which expressed mC herry‐Cre fusion protein. Both of the fluorescence images from mC herry and Cre induced mC herry in a reporter rat strain, showing that ISCA 1 expressed in most of the tissues in rats. The ISCA 1 knockout resulted in abnormal development at 8.5 days, with a significant decrease of NDUFA 9 protein and an increase of aconitase 2 ( ACO 2) in rat embryos. Conclusion Deletion of ISCA 1 induced early death in rats. ISCA 1 affected the expression of key proteins in the mitochondrial respiratory chain complex, suggesting that ISCA 1 has an important influence on the respiratory complex and energy metabolism.