Facultad de Agronomía, Departamento de Producción Animal, Buenos Aires, Laboratorio Biotecnología Animal (LabBA), Universidad de Buenos Aires, Buenos Aires, Argentina.
Instituto de Investigaciones en Producción Animal (INPA), CONICET-Universidad de Buenos Aires, Buenos Aires, Argentina.
Methods Mol Biol. 2022;2495:129-148. doi: 10.1007/978-1-0716-2301-5_8.
The CRISPR-on system is a programmable, simple, and versatile gene activator that has proven to be efficient in cultured cells from several species and in bovine embryos. This technology allows for the precise and specific activation of single endogenous gene expression and also multiplexed gene expression in a simple fashion. Therefore, CRISPR-on has unique advantages over other activator systems and a wide adaptability for studies in basic and applied science, such as cell reprogramming and cell fate differentiation for regenerative medicine.In this chapter, we describe the materials and methods of the CRISPR-on system for activation of the endogenous SMARCA4 expression in bovine embryos.
CRISPR-on 系统是一种可编程、简单且多功能的基因激活系统,已被证明在来自多个物种的培养细胞和牛胚胎中具有高效性。该技术允许以简单的方式精确和特异性地激活单个内源性基因表达,也可以实现多基因表达的多路复用。因此,CRISPR-on 相对于其他激活系统具有独特的优势,并且在基础科学和应用科学研究中具有广泛的适应性,例如细胞重编程和细胞命运分化用于再生医学。在本章中,我们描述了用于激活牛胚胎中内源性 SMARCA4 表达的 CRISPR-on 系统的材料和方法。