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Article

CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori

by
Hao Guo
1,2,3,
Feng Chen
1,2,3,
Mingyi Zhou
1,2,3,
Weiqun Lan
1,2,3,
Wenchang Zhang
1,2,3,
Guanwang Shen
1,2,3,
Ping Lin
1,2,3,
Qingyou Xia
1,2,3,
Ping Zhao
1,2,3 and
Zhiqing Li
1,2,3,*
1
State Key Laboratory of Silkworm Genome Biology, Biological Science Research Center, Southwest University, Chongqing 400715, China
2
Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City & Southwest University, Chongqing 400715, China
3
Chongqing Key Laboratory of Sericultural Science, Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Chongqing 400715, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2023, 24(4), 3468; https://doi.org/10.3390/ijms24043468
Submission received: 22 December 2022 / Revised: 1 February 2023 / Accepted: 7 February 2023 / Published: 9 February 2023
(This article belongs to the Special Issue Epigenetic Regulation of Gene Expression)
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Abstract

DNA N6-methyladenine (6mA) has recently been found to play regulatory roles in gene expression that links to various biological processes in eukaryotic species. The functional identification of 6mA methyltransferase will be important for understanding the underlying molecular mechanism of epigenetic 6mA methylation. It has been reported that the methyltransferase METTL4 can catalyze the methylation of 6mA; however, the function of METTL4 remains largely unknown. In this study, we aim to investigate the role of the Bombyx mori homolog METTL4 (BmMETTL4) in silkworm, a lepidopteran model insect. By using CRISPR-Cas9 system, we somatically mutated BmMETTL4 in silkworm individuates and found that disruption of BmMETTL4 caused the developmental defect of late silkworm embryo and subsequent lethality. We performed RNA-Seq and identified that there were 3192 differentially expressed genes in BmMETTL4 mutant including 1743 up-regulated and 1449 down-regulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that genes involved in molecular structure, chitin binding, and serine hydrolase activity were significantly affected by BmMETTL4 mutation. We further found that the expression of cuticular protein genes and collagens were clearly decreased while collagenases were highly increased, which had great contributions to the abnormal embryo and decreased hatchability of silkworm. Taken together, these results demonstrated a critical role of 6mA methyltransferase BmMETTL4 in regulating embryonic development of silkworm.
Keywords: Bombyx mori; CRISPR-Cas9; METTL4; N6-methyladenine; embryonic development Bombyx mori; CRISPR-Cas9; METTL4; N6-methyladenine; embryonic development

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MDPI and ACS Style

Guo, H.; Chen, F.; Zhou, M.; Lan, W.; Zhang, W.; Shen, G.; Lin, P.; Xia, Q.; Zhao, P.; Li, Z. CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori. Int. J. Mol. Sci. 2023, 24, 3468. https://doi.org/10.3390/ijms24043468

AMA Style

Guo H, Chen F, Zhou M, Lan W, Zhang W, Shen G, Lin P, Xia Q, Zhao P, Li Z. CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori. International Journal of Molecular Sciences. 2023; 24(4):3468. https://doi.org/10.3390/ijms24043468

Chicago/Turabian Style

Guo, Hao, Feng Chen, Mingyi Zhou, Weiqun Lan, Wenchang Zhang, Guanwang Shen, Ping Lin, Qingyou Xia, Ping Zhao, and Zhiqing Li. 2023. "CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori" International Journal of Molecular Sciences 24, no. 4: 3468. https://doi.org/10.3390/ijms24043468

APA Style

Guo, H., Chen, F., Zhou, M., Lan, W., Zhang, W., Shen, G., Lin, P., Xia, Q., Zhao, P., & Li, Z. (2023). CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori. International Journal of Molecular Sciences, 24(4), 3468. https://doi.org/10.3390/ijms24043468

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