Generation of prolactin-inducible protein (Pip) knockout mice by CRISPR/Cas9-mediated gene engineering
| Autor: | Lucas E. L. Terceiro, Chidalu A Edechi, Anne Blanchard, Yvonne Myal, Barbara Triggs-Raine, Etienne Leygue, Agnes Fresnoza |
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| Rok vydání: | 2022 |
| Předmět: |
Mice
Knockout musculoskeletal diseases Pharmacology chemistry.chemical_classification Physiology Apocrine Proteins General Medicine Biology Gene engineering Molecular biology Mice Inbred C57BL Gene Knockout Techniques Prolactin-Inducible Protein chemistry CRISPR-Associated Protein 9 Physiology (medical) Models Animal Knockout mouse Animals CRISPR lipids (amino acids peptides and proteins) CRISPR-Cas Systems Genetic Engineering Glycoprotein |
| Zdroj: | Canadian Journal of Physiology and Pharmacology. 100:86-91 |
| ISSN: | 1205-7541 0008-4212 |
| DOI: | 10.1139/cjpp-2021-0306 |
| Popis: | Prolactin-inducible protein (PIP) is a multifunctional glycoprotein that is highly expressed and found in the secretions of apocrine glands such as salivary, lacrimal, and sweat glands including the mammary glands. PIP has been implicated in various diseases, including breast cancer, gross cystic disease of the breast, keratoconus of the eye, and the autoimmune Sjögren’s syndrome. Here we have generated a Pip knockout (KO) mouse using the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRSPR-associated (Cas)9 system. The Cas9 protein and two single guide RNAs targeting specific regions for both exons 1 and 2 of the Pip gene were microinjected into mouse embryos. The deletions and insertions promoted by CRISPR/Cas9 system on the Pip gene successfully disrupted Pip protein coding, as confirmed by PCR genotyping, sequencing, and ultimately Western blot analysis. This mouse model was generated in the inbred C57Bl/6J mouse, which exhibits lower genetic variation. This novel CRISPR Pip KO mouse model will not only be useful for future studies to interrogate the multifunctional role of PIP in physiological processes but will facilitate a broader understanding of the function of PIP in vivo while providing unprecedented insight into its role in a spectrum of diseases attributed to the deregulation of the PIP gene. |
| Databáze: | OpenAIRE |
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