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Updated: Nov 24, 2025
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
Published on: November 1, 2024
Generation of a MCPH1 knockout human embryonic stem cell line by CRISPR/Cas9 technology
Zerui Wang1, Yazhou Cui1, Yongli Shan2
1Department of Orthopedic Surgery, The First Affiliated Hospital of Shandong First Medical University, Ji'nan 250014, Shandong, China; Biomedical Sciences College & Shandong Medicinal Biotechnology Centre, Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan 250062, Shandong, China; Key Lab for Biotech-Drugs of National Health Commission, Ji'nan 250062, Shandong, China; Key Lab for Rare & Uncommon Diseases of Shandong Province, Ji'nan 250062, Shandong, China.
Abstract:
Human MCPH1 (Microcephalin 1) encodes a DNA damage response protein. Mutations in this gene have been associated with Primary Autosomal Recessive Microcephaly and premature chromosome condensation syndrome. To further understand the roles of MCPH1 in neural differentiation and brain development, here we generated a MCPH1 knockout human embryonic stem cell line by CRISPR/Cas9 genome editing technology. This cell line maintained a normal karyotype and typical undifferentiated state in terms of morphology, pluripotent gene expression, and had differentiation potential in vitro. This cell line provides a good resource to study the role of MCPH1 gene in neurogenesis and regulation of the size of the cerebral cortex in vitro.
