CRISPR/Cas9-mediated gene editing on Sox2ot promoter leads to its truncated expression and does not influence neural tube closure and embryonic development in mice
| Autor: | San-Qiang Li, She-Gan Gao, Dao-Yin Dong, Pu-Yu Li |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Gene Editing
Neural Tube Embryogenesis Biophysics Neural tube Embryonic Development Cell Biology Biology Biochemistry Cell biology Neuroepithelial cell Mice Exon medicine.anatomical_structure SOX2 Gene expression medicine Animals RNA Long Noncoding Guide RNA CRISPR-Cas Systems Promoter Regions Genetic Molecular Biology Gene |
| Zdroj: | Biochemical and Biophysical Research Communications. 573:107-111 |
| ISSN: | 0006-291X |
| Popis: | Sox2 overlapping transcript (Sox2ot) is a long non-coding RNA (lncRNA), which harbors one of the major regulators of pluripotency, the Sox2 gene, in its intronic region. Sox2ot is primarily expressed in the developing neuroepithelium. However, its role in neural tube closure and embryonic development remains unclear. To investigate if Sox2ot is required for neural tube closure and embryonic development, Sox2ot promoter was deleted by CRISPR-Cas9 genome editing technology to prevent Sox2ot gene expression in mice. We designed 9 guide RNAs to specifically target the Sox2ot promoter and 3 gRNAs induced gene editing on the promoter of the Sox2ot gene in cells transfected with Cas9 mRNA and gRNAs. Then, these gRNAs and Cas9 mRNA were injected into mouse zygotes and implanted into pseudopregnant mice. A Sox2ot promoter-deleted mouse line was identified with complete deletion of promoter as well as deletion of exon 1 and exon 2. Sox2ot transcript was truncated with a lack of exon 1 and exon 2 in Sox2ot promoter-deleted mice. Furthermore, neural tube closure and embryonic development were checked at E9.5, E10.5, E14.5, E17.5 and after-birth (P2) and we did not find any failure of neural tube closure and aberrant embryonic development in Sox2ot promoter-deleted mice. Thus, our study demonstrated that CRISPR-Cas9 gene editing in Sox2ot promoter leads to its truncated expression and does not influence neural tube closure and embryonic development. |
| Databáze: | OpenAIRE |
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