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Application of Mouse Parthenogenetic Haploid Embryonic Stem Cells as a Substitute of Sperm

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Published on: November 19, 2020

Genetic modification and screening in rat using haploid embryonic stem cells.

Wei Li¹, Xin Li², Tianda Li¹

¹State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.

|December 24, 2013

Summary

Researchers developed rat androgenetic haploid embryonic stem cells (RahESCs) to overcome genetic manipulation challenges in rat models. These RahESCs enable efficient homozygous mutation generation and the creation of transgenic rat lines for biomedical research.

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Area of Science:

Biomedical Research
Developmental Biology
Genetics

Background:

Rats are crucial animal models in biomedical research.
Genetic manipulation in rats faces practical limitations, hindering genetic analysis.
A need exists for advanced tools to facilitate genetic studies in rats.

Purpose of the Study:

To derive rat androgenetic haploid embryonic stem cells (RahESCs).
To establish RahESCs as a tool for genetic analysis and manipulation in rats.
To enable the production of genetically modified rats.

Main Methods:

Removal of the maternal pronucleus from rat zygotes to create androgenetic embryos.
Derivation of embryonic stem cells (ESCs) from these embryos.
Utilizing gene trapping, homologous recombination, and CRISPR-Cas for gene modification.
Intracytoplasmic injection of RahESCs into oocytes.

Main Results:

Successfully derived rat androgenetic haploid embryonic stem cells (RahESCs).
RahESCs express pluripotency markers, differentiate into three germ layers, and contribute to the germline.
Homozygous mutations were introduced using gene trapping and gene targeting.
Fertile rats were produced from RahESCs, transmitting genetic modifications to offspring.

Conclusions:

RahESCs provide a practical solution for genetic manipulation in rats.
This technology facilitates functional genetic studies and the generation of transgenic rat lines.
RahESCs significantly advance the utility of rats in biomedical research.