LEIi004-A-1

Not yet reviewed by Australian Stem Cell Registry

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)
retinitis pigmentosa (Variant) retinitis pigmentosa (Variant)

Genetic Information

Genotype Locus
CLN3
Polymorphism
Heterozygous: NM_000086.2:c.175G=(p.Ala59=); Heterozygous: unspecified_reference_CLN3: 1.02kb deletion including of exons 7 and 8
Modifications
Variant Variant Isogenic modification

Associated Publications

Journal Year Article
Molecules (Basel, Switzerland) 2021 Patient-Derived Induced Pluripotent Stem Cell Models for Phenotypic Screening in the Neuronal Ceroid Lipofuscinoses
Molecular genetics & genomic medicine 2021 Gene correction of the CLN3 c.175G>A variant in patient-derived induced pluripotent stem cells prevents pathological changes in retinal organoids
Molecular Genetics and Genomic Medicine 2021 Gene correction of the CLN3 c.175G>A variant in patient-derived induced pluripotent stem cells prevents pathological changes in retinal organoids.
Stem cell research 2018 Generation of an induced pluripotent stem cell line from a patient with non-syndromic CLN3-associated retinal degeneration and a coisogenic control line
Stem Cell Research 2018 Generation of an induced pluripotent stem cell line from a patient with non-syndromic CLN3-associated retinal degeneration and a coisogenic control line.

Line Custodianship

Cell Line Maintainer

Chen

Affiliated Institutions
  • Lions Eye Institute, Nedlands, Australia
View all cell lines from this group

Cell Line Producer

Lions Eye Institute

Affiliated Institutions
  • Lions Eye Institute, Nedlands, Australia

Derivation Details

Induced Pluripotent Cell Derivation Details

Source Cell Type: CL:0000057
Label: fibroblast
Definition: A connective tissue cell which secretes an extracellular matrix rich in collagen and other macromolecules. Flattened and irregular in outline with branching processes; appear fusiform or spindle-shaped.

Source Cell Origin: N/A

Derivation Year:

Reprogramming Method

Vector Type: Not integrated

Vector: Episomal

Ethics

Ethics Number: 2001–053

Institution Human Research Ethics Council: Sir Charles Gairdner Hospital Human Research Ethics Committee

Approval Date: To be verified

Modifications

Genomic Modifications

Retinitis pigmentosa (RP) is an inherited retinal dystrophy leading to progressive loss of the photoreceptors and retinal pigment epithelium and resulting in blindness usually after several decades.

Mutation Type: Variant

Retinitis pigmentosa (RP) is an inherited retinal dystrophy leading to progressive loss of the photoreceptors and retinal pigment epithelium and resulting in blindness usually after several decades.

Mutation Type: Variant

Heterozygous: NM_000086.2:c.175G=(p.Ala59=); Heterozygous: unspecified_reference_CLN3: 1.02kb deletion including of exons 7 and 8

Correction of heterozygous NM_000086.2 (CLN3):c.175G>A(p.Ala59Thr) present in the parental line (LEIi004-A). Note that the 1.02kb deletion present in the parental line is retained (Heterozygous: unspecified_reference_CLN3: 1.02kb deletion including of exons 7 and 8). Correction was confirmed by PCR and Sanger sequencing, and screening for off-target modifications ensured no additional variations were introduced.

Cytoband: 16p12.1
Mutation Type: Isogenic modification
Delivery Method: CRISPR/Cas9

Quality Assurance

Genomic Characterisation

Passage Number: 41

Karyotype: arr(1-22,X)x2

Karyotype Method: Molecular karyotyping by SNP array

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

STR analysis Results:

Loci Allele 1 Allele 2
CSF1PO None None
D13S317 None None
D16S539 None None
D18S51 None None
D21S11 None None
D3S1358 None None
D5S818 None None
D7S820 None None
D8S1179 None None
FGA None None
PENTA D None None
PENTA E None None
TH01 None None
TPOX None None
VWA None None
Amelogenin None None

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
KLF4 Immunostaining
NANOG Immunostaining
SOX2 Immunostaining
SSEA4 Immunostaining
TRA-1-81 Immunostaining
C-MYC RT-PCR
KLF4 RT-PCR
SOX2 RT-PCR
NANOG RT-PCR
POU5F1 (OCT-4) Immunostaining
POU5F1 (OCT-4) RT-PCR
SSEA-4 Immunostaining
TRA 1-81 Immunostaining

Scorecard Results

Undifferentiated Cells

No available data for this cell line.

Pluripotency

No available data for this cell line.

Pluripotency Characterisation

Endoderm

In vitro spontaneous differentiation

Assessed by: RT-PCR

Markers: Brachyury;FDGFRB

Mesoderm

In vitro spontaneous differentiation

Assessed by: RT-PCR

Markers: FOXA2;GATA4;SOX17

Ectoderm

In vitro spontaneous differentiation

Assessed by: RT-PCR

Markers: OTX1;PAX6

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: Unavailable

O2 concentration: Unavailable

Medium Items:

  • Base Medium: TeSR-E8 - STEMCELL Technologies
  • Base Coat: Geltrex - ThermoFisher

Passage Method: Enzyme-free cell dissociation

External References

Source
Molecules (Basel, Switzerland) Patient-Derived Induced Pluripotent Stem Cell Models for Phenotypic Screening in the Neuronal Ceroid Lipofuscinoses
Molecular genetics & genomic medicine Gene correction of the CLN3 c.175G>A variant in patient-derived induced pluripotent stem cells prevents pathological changes in retinal organoids
Molecular Genetics and Genomic Medicine Gene correction of the CLN3 c.175G>A variant in patient-derived induced pluripotent stem cells prevents pathological changes in retinal organoids.
Stem cell research Generation of an induced pluripotent stem cell line from a patient with non-syndromic CLN3-associated retinal degeneration and a coisogenic control line
Stem Cell Research Generation of an induced pluripotent stem cell line from a patient with non-syndromic CLN3-associated retinal degeneration and a coisogenic control line.
hPSCreg LEIi004-A-1
Cellosaurus CVCL_VE94
BioSamples SAMEA4760675
Wikidata Q54902438
Phenomics Australia Phenomics Australia