Oocyte-specific gene knockdown by intronic artificial microRNAs driven by Zp3 transcription in mice.

Keisuke Sasaki, Saaya Takaoka, Yayoi Obata

The Journal of reproduction and development 2021 Jun 21

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Conditional knockout technology is a powerful tool for investigating the spatiotemporal functions of target genes. However, generation of conditional knockout mice involves complicated breeding programs and considerable time. A recent study has shown that artificially designed microRNAs (amiRNAs), inserted into an intron of the constitutively expressed gene, induce knockdown of the targeted gene in mice, thus creating a simpler method to analyze the functions of target genes in oocytes. Here, to establish an oocyte-specific knockdown system, amiRNA sequences against enhanced green fluorescent protein (EGFP) were knocked into the intronic sites of the Zp3 gene. Knock-in mice were then bred with EGFP transgenic mice. Our results showed that Zp3-derived amiRNA successfully reduced EGFP fluorescence in the oocytes in a size-dependent manner. Importantly, knockdown of EGFP did not occur in somatic cells. Thus, we present our knockdown system as a tool for screening gene functions in mouse oocytes.

Citation

Keisuke Sasaki, Saaya Takaoka, Yayoi Obata. Oocyte-specific gene knockdown by intronic artificial microRNAs driven by Zp3 transcription in mice. The Journal of reproduction and development. 2021 Jun 21;67(3):229-234