Pronuclear Microinjection of One-Cell Embryos

Area of Science:

• Developmental Biology
• Genetics
• Molecular Biology

Background:

• Transgenic and gene-modified mice are crucial models in biological research.
• Generating these models requires introducing exogenous genetic material into early-stage embryos.
• The mouse zygote, containing two pronuclei, is the target for these genetic modifications.

Purpose of the Study:

• To present detailed procedures for pronuclear microinjection in mouse zygotes.
• To provide a foundational technique applicable to various genetic engineering projects.

Main Methods:

• Microinjection of DNA constructs into the male pronucleus of mouse zygotes.
• Application of the technique for generating traditional transgenic mice.
• Adaptation of the method for introducing gene-editing reagents (ZFNs, TALENs, CRISPR RNAs).

Main Results:

• Successful introduction of exogenous reagents into mouse zygotes via pronuclear microinjection.
• Demonstration of the technique's versatility for diverse genetic modification goals.

Conclusions:

• Pronuclear microinjection is a fundamental and widely applicable technique for creating genetically modified mice.
• This method facilitates advancements in both transgenesis and precise gene editing in research models.