Generation of a FRMD5 knockout human embryonic stem cell line by CRISPR/Cas9 editing

Area of Science:

• Genetics
• Neuroscience
• Developmental Biology

Background:

• FRMD5 is a gene on chromosome 15q15.3 encoding a FERM domain protein.
• Mutations in FRMD5 are associated with NEDEMA, a rare neurodevelopmental disorder.
• NEDEMA presents with developmental delay, intellectual disability, ataxia, epilepsy, and abnormal eye movements.

Purpose of the Study:

• To establish a human embryonic stem cell (hESC) model for FRMD5.
• To create a tool for investigating the role of FRMD5 in neural development.

Main Methods:

• CRISPR/Cas9 gene editing was used to create the FRMD5 knockout hESC line.
• Standard cell culture techniques were employed to maintain pluripotency and assess differentiation potential.

Main Results:

• A FRMD5 knockout (FRMD5^-/^-) hESC line was successfully established.
• The FRMD5^-/^- hESC line maintained a normal karyotype.
• The cell line expressed key pluripotency markers and differentiated into all three germ layers.

Conclusions:

• The FRMD5^-/^- hESC line is a viable model for studying FRMD5 function.
• This model will facilitate research into the genetic basis of NEDEMA and FRMD5-related neurodevelopmental disorders.