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Updated: May 12, 2026
CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models
Published on: August 24, 2022
A new chemical approach to the efficient generation of mouse embryonic stem cells
Hossein Baharvand1, Seyedeh-Nafiseh Hassani
1Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Abstract:
Here, we present a highly efficient and reproducible method for the establishment of mouse embryonic stem cells (mESCs) from embryonic day 3.5 (E3.5) whole blastocysts. This protocol involves the use of small molecules SB431542 and PD0325901, which inhibit transforming growth factor-β (TGF-β) and extracellular signal-regulated kinases (ERK1/2), respectively. This protocol is universal in the derivation of mESC lines from NMRI, C57BL/6, BALB/c, DBA/2, and FVB/N strains, which have previously been considered refractory or non-permissive for ESC establishment. The efficiency of mESC lines generation is 100%, regardless of genetic background.
