WAe009-A-3C

Flagged for further review

Alternative Names: AGK-KO C10; AGKKO C10

Synonyms: MCRIe010-A-3C

Details

Description

This embryonic stem cell line is a subclone of WAe009-A which was originally derived from a female blastocyst. WAe009-A-3C has been modified using CRISPR/Cas9 to induce biallelic knockout of the acylglycerol kinase (AGK) gene via compound heterozygous frame shifting mutations. The line represents a model of the mitochondrial disease Sengers syndrome. Expression of OCT4, NANOG, SSEA-4, TRA-1-60 and TRA-1-81 in undifferentiated cells, and directed differentiation to endoderm, mesoderm and ectoderm have been reported.

Tissue & Disease as reported

Disease as reported (Genomic Modifications)

Genetic Information

Genotype Locus
AGK
Polymorphism
Compound heterozygous: NC_000007.14:g.141596617_141596618del; and: NC_000007.14:g.141596619insT
Modifications
Gene Knock-out

Associated Publications

Journal Year Article
Stem Cell Research 2026 Generation of a pluripotent human AGK knockout embryonic stem cell model (WAe009-A-3C) of Sengers syndrome

Line Custodianship

Cell Line Maintainer

Ann Frazier

Affiliated Institutions
  • Murdoch Children's Research Institute, Melbourne, Australia
View all cell lines from this group

Cell Line Producer

Murdoch Children's Research Institute

Affiliated Institutions
  • Murdoch Children's Research Institute, Melbourne, Australia

Source

Parental Cell Line

WAe009-A

Donor Derived

Age: Embryo

Biological Sex: Female

Derivation Details

Embryonic Derivation Details

NHMRC Licence Number: None

Genetic Diagnosis: None

Supernumerary from IVF: None

Derivation Year: None

Ethics

Ethics Number: 93025

Institution Human Research Ethics Council: Royal Children's Hospital Human Research Ethics Committee

Approval Date: None

Modifications

Genomic Modifications

AGK

CRISPR/Cas9 was used introduce frame-shifting compound heterozygous mutations (2 bp deletion, 1 bp insertion) into exon four of the AGK gene. Alleles were confirmed by PCR and Sanger sequencing and loss of protein expression was confirmed by Western Blot. Absence of plasmid integration was confirmed by PCR. The top five predicted off-target sites were PCR amplified and Sanger sequenced, and no off-target effects were observed.

Genotype: Heterozygous: NC_000007.14:g.141596619insT
Cytoband: 7q34
Mutation Type: Gene Knock-out
Delivery Method: CRISPR/Cas9

Quality Assurance

Genomic Characterisation

Passage Number: Not available

Karyotype: arr(1-22,X)x2

Karyotype Method: Molecular karyotyping by SNP array

Summary: Molecular karyotyping by SNP array (Illumina HumanCoreExome-24 v 1.1; resolution 0.5 Mb).

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
NANOG Immunostaining
POU5F1 (OCT-4) Flow cytometry
SSEA-4 Flow cytometry
TRA-1-81 Flow cytometry
TRA-1-60 Flow cytometry

Scorecard Results

Undifferentiated Cells

No available data for this cell line.

Pluripotency

No available data for this cell line.

Pluripotency Characterisation

Endoderm

In vitro directed differentiation

Assessed by: In vitro directed differentiation

Markers: SOX17, CXCR4

Mesoderm

In vitro directed differentiation

Assessed by: In vitro directed differentiation

Markers: TBXT, CXCR4

Ectoderm

In vitro directed differentiation

Assessed by: In vitro directed differentiation

Markers: PAX6, NES

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: 5%

O2 concentration: 5%

Medium Items:

  • Base Medium: DMEM F/12
  • Serum: KnockOut Serum Replacement - Thermo Fisher Scientific Inc.
  • Supplement: FGF2 - PeproTech

Passage Method: Enzymatically