hPSCreg®
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EDi001-A-3

Registration Summary:

AST23_SNCAKO Clone 1, AST22-1KO-1, AST23-1KO-1, AST22_SNCAKO Clone 1

EDi001-A-3

General

Cell Line
hPSCreg Name
EDi001-A-3
Alternative name(s)
AST23_SNCAKO Clone 1, AST22-1KO-1, AST23-1KO-1, AST22_SNCAKO Clone 1
Cell line type Human induced pluripotent stem cell (hiPSC)
Similar lines
Last update 14th May 2022
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Provider
Generator University of Edinburgh (ED)
Owner College of Medicine and Veterinary Medicine
Distributors
Derivation country United States

External Databases
BioSamples SAMEA3323899
Cellosaurus CVCL_LE53
ECACC 66540168
EBiSC EDi001-A-3
CLO CLO_0102714
Wikidata Q54831974

General Information
Projects
* Is the cell line readily obtainable for third parties?
Yes
Research use: allowed
Clinical use: allowed
Commercial use: allowed
Subclone of

Donor Information

General Donor Information
Sex female

Phenotype and Disease related information (Donor)
Diseases A disease was diagnosed.
Parkinson disease
Parkinson disease in the OLS
This is a PD line, with the control being EDi002-A lines and CRISPR/Cas9-corrected EDi001-A-1, EDi001-A-2, EDi001-A-3 and EDi001-A-4
The donor is a carrier of a disease-associated mutation and affected.
Genetic variants
SNCA (target)
4q22.1
Heterozygous
The donor carries a triplication of the alpha-synuclein gene, resulting in 4 copies of SNCA. The copies of SNCA are situated in a heterozygous triplication configuration. See Figure 1 of Petrucci, 2015 for a graphic representation of the heterozygous triplication.
Family history Strong family history of Parkinson’s disease due to autosomal dominant inheritance of SNCA triplication
Is the medical history available upon request? Y Mov Disord. 2011 Sep;26(11):2134-6. doi: 10.1002/mds.23776

Donor Relations
Other cell lines of this donor
All cell lines of this donor's relatives
Has daughter:

External Databases (Donor)
BioSamples SAMEA3319991

Ethics

Also have a look at the ethics information for the parental line EDi001-A .
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used?

hIPSC Derivation

General
The source cell information can be found in the parental cell line EDi001-A.

Reprogramming method
Vector type Integrating
Vector Virus (Retrovirus)
Genes
Is the used vector excisable?
No
Absence of reprogramming vector(s)?
Unknown
Reprogramming vectors silenced?
Yes
Methods used
RT-PCR

Vector free reprogramming
Type of used vector free reprogramming factor(s)
None

Other
Selection criteria for clones Nickase pair mediated SNCA Knock-out of 1 SNCA alleles. Therefore 3 alleles remain.
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions

Surface coating Laminin
Feeder cells
No
Passage method Enzymatically
Accutase
O2 Concentration 95 %
CO2 Concentration 5 %
Medium Essential 8™
Supplements
Rock inhibitor added while passaging 10 nM

Characterisation

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
SSEA-4
Yes
TRA 1-60
Yes
SSEA-1
No
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
Mesoderm
Ont Id: UBERON_0000926
Ectoderm
Ont Id: UBERON_0000924

Microbiology / Virus Screening
HIV 1 Negative
HIV 2 Negative
Hepatitis B Negative
Hepatitis C Negative
Mycoplasma Negative

Certificate of Analysis
Is there a certificate of analysis available?
Yes
EDi001-A-3.P001_.CoA_.v1_.pdf
Passage: 58

Genotyping

Karyotyping (Cell Line)
Has the cell line karyotype been analysed?
No

Other Genotyping (Cell Line)

Genetic Modification

Disease/phenotype related modifications
Parkinson disease
Parkinson disease in the OLS
Genetic modifications
SNCA (target)
Gene knock-out
4q22.1
CRISPR/Cas was employed to create a mutation in Exon2, disrupting the ATG start site and some sequence 5' to the coding start of SNCA. PCR amplicons of the CRISPR site were cloned using the TOPO cloning technique. Sequencing of 8 TOPO clones detected 3 deletions; however, sequencing of the CRISPR sites shows that only one of these modifications disrupts the ATG site. This indicates 1 allele was putatively knocked out, and 3 alleles remain. Additional comment: there are two additional ATG sites located immediately downstream of the normal ATG start site, one of which is in-frame (9 bases) to the initial ATG. It is possible that inactivation of the first ATG start site might not be completely sufficient to prevent translation from the second downstream ATG, which is in-frame to the first.
CRISPR-associated (CRISPR/Cas) System