CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways

Ganglioside-induced differentiation-associated protein 2 (<i>GDAP2</i>) is a gene involved in hereditary cerebellar ataxia. At present, little is known about the function of <i>GDAP2</i> in insects. In this study, <i>BmGDAP2</i> was detected to be highly expressed...

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Main Authors: Chaojun Yuan, Zichong Zhou, Qifeng Guo, Ying Yang, Yue Sun, Yong Liu, Wenyi Jia, Shuoqi Fan, Jinxin Wu, Xiaoting Hua, Ping Lin, Ping Zhao, Qingyou Xia
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Insects
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Online Access:https://www.mdpi.com/2075-4450/16/4/354
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author Chaojun Yuan
Zichong Zhou
Qifeng Guo
Ying Yang
Yue Sun
Yong Liu
Wenyi Jia
Shuoqi Fan
Jinxin Wu
Xiaoting Hua
Ping Lin
Ping Zhao
Qingyou Xia
author_facet Chaojun Yuan
Zichong Zhou
Qifeng Guo
Ying Yang
Yue Sun
Yong Liu
Wenyi Jia
Shuoqi Fan
Jinxin Wu
Xiaoting Hua
Ping Lin
Ping Zhao
Qingyou Xia
author_sort Chaojun Yuan
collection DOAJ
description Ganglioside-induced differentiation-associated protein 2 (<i>GDAP2</i>) is a gene involved in hereditary cerebellar ataxia. At present, little is known about the function of <i>GDAP2</i> in insects. In this study, <i>BmGDAP2</i> was detected to be highly expressed in the head, epidermis, midgut, and anterior silk glands of silkworms. We generated a knockout mutant, <i>BmGDAP2</i> (<i>BmGDAP2</i><sup>KO</sup>), using the CRISPR/Cas9 system. Compared with that of the wild-type, the growth cycle of <i>BmGDAP2</i><sup>KO</sup> larvae was significantly prolonged, while their body size was reduced. Furthermore, we found 149 differentially expressed genes (DEGs) between <i>BmGDAP2</i><sup>KO</sup> and the wild-type, including 106 upregulated and 43 downregulated genes. GO annotation analysis indicated that <i>BmGDAP2</i> primarily influences structural and molecular activities, as well as catalytic and binding functions. KEGG pathway analysis revealed that the differentially expressed genes were mainly enriched in pathways related to peroxidase activity, hormone synthesis, apoptosis, and longevity regulation. Further investigation focused on candidate genes related to these pathways. We found that the expression levels of <i>MAD2L1</i>, which can inhibit cell proliferation and promote apoptosis, and <i>Aurka</i>-b, which plays a crucial role in cell cycle regulation, were significantly reduced in <i>BmGDAP2</i><sup>KO</sup> silkworms. These changes may interfere with the normal functions of cell division, leading to the prolonged developmental cycle observed in <i>BmGDAP2</i><sup>KO</sup> larvae. Our findings demonstrate that knockout of <i>BmGDAP2</i> significantly prolongs the life cycle of <i>Bombyx mori</i> by affecting genes related to autophagy, apoptosis, and hormone regulation.
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spelling doaj-art-5fa7708311ae4ff4a3daa64e8932ae682025-08-20T02:28:36ZengMDPI AGInsects2075-44502025-03-0116435410.3390/insects16040354CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental PathwaysChaojun Yuan0Zichong Zhou1Qifeng Guo2Ying Yang3Yue Sun4Yong Liu5Wenyi Jia6Shuoqi Fan7Jinxin Wu8Xiaoting Hua9Ping Lin10Ping Zhao11Qingyou Xia12Integrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City, Biological Science Research Center, Southwest University, Chongqing 400715, ChinaGanglioside-induced differentiation-associated protein 2 (<i>GDAP2</i>) is a gene involved in hereditary cerebellar ataxia. At present, little is known about the function of <i>GDAP2</i> in insects. In this study, <i>BmGDAP2</i> was detected to be highly expressed in the head, epidermis, midgut, and anterior silk glands of silkworms. We generated a knockout mutant, <i>BmGDAP2</i> (<i>BmGDAP2</i><sup>KO</sup>), using the CRISPR/Cas9 system. Compared with that of the wild-type, the growth cycle of <i>BmGDAP2</i><sup>KO</sup> larvae was significantly prolonged, while their body size was reduced. Furthermore, we found 149 differentially expressed genes (DEGs) between <i>BmGDAP2</i><sup>KO</sup> and the wild-type, including 106 upregulated and 43 downregulated genes. GO annotation analysis indicated that <i>BmGDAP2</i> primarily influences structural and molecular activities, as well as catalytic and binding functions. KEGG pathway analysis revealed that the differentially expressed genes were mainly enriched in pathways related to peroxidase activity, hormone synthesis, apoptosis, and longevity regulation. Further investigation focused on candidate genes related to these pathways. We found that the expression levels of <i>MAD2L1</i>, which can inhibit cell proliferation and promote apoptosis, and <i>Aurka</i>-b, which plays a crucial role in cell cycle regulation, were significantly reduced in <i>BmGDAP2</i><sup>KO</sup> silkworms. These changes may interfere with the normal functions of cell division, leading to the prolonged developmental cycle observed in <i>BmGDAP2</i><sup>KO</sup> larvae. Our findings demonstrate that knockout of <i>BmGDAP2</i> significantly prolongs the life cycle of <i>Bombyx mori</i> by affecting genes related to autophagy, apoptosis, and hormone regulation.https://www.mdpi.com/2075-4450/16/4/354<i>GDAP2</i>silkwormdevelopmentlifespan
spellingShingle Chaojun Yuan
Zichong Zhou
Qifeng Guo
Ying Yang
Yue Sun
Yong Liu
Wenyi Jia
Shuoqi Fan
Jinxin Wu
Xiaoting Hua
Ping Lin
Ping Zhao
Qingyou Xia
CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
Insects
<i>GDAP2</i>
silkworm
development
lifespan
title CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
title_full CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
title_fullStr CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
title_full_unstemmed CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
title_short CRISPR/Cas9-Mediated Knockout of <i>BmGDAP2</i> in the Silkworm, <i>Bombyx mori</i>: Extended Lifespan and Altered Gene Expression Impacting Developmental Pathways
title_sort crispr cas9 mediated knockout of i bmgdap2 i in the silkworm i bombyx mori i extended lifespan and altered gene expression impacting developmental pathways
topic <i>GDAP2</i>
silkworm
development
lifespan
url https://www.mdpi.com/2075-4450/16/4/354
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