AST22-2KO-6, AST23_SNCAKO Clone 6, AST22_SNCAKO Clone 6, AST23-2KO-6

General#

Cell Line

hPSCreg Name EDi001-A-4
Alternative name(s)
AST22-2KO-6, AST23_SNCAKO Clone 6, AST22_SNCAKO Clone 6, AST23-2KO-6
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 2nd July 2019
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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 SAMEA3323960
Cellosaurus CVCL_LE54
ECACC 66540169
EBiSC EDi001-A-4

General Information

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

Donor Information#

General Donor Information

Sex female

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
Parkinson's disease
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 affected.
Synonyms
  • Parkinson's syndrome
  • Parkinsons
  • Primary Parkinsonism
  • Parkinsons disease
  • Parkinson disease
  • Parkinson's disease (disorder)
  • Parkinson's disease NOS
  • Parkinson Disease, Idiopathic
  • PARKINSON DIS
  • Paralysis agitans
  • IDIOPATHIC PARKINSONS DIS
  • PARKINSON DIS IDIOPATHIC
  • Parkinsonism, Primary
  • Parkinson's Disease, Lewy Body
  • IDIOPATHIC PARKINSON DIS
  • Idiopathic PD
  • Idiopathic Parkinson Disease
  • Lewy Body Parkinson's Disease
  • Parkinsonian disorder
  • LEWY BODY PARKINSON DIS
  • Parkinson's
  • Idiopathic Parkinson's Disease
  • Parkinson's disease NOS (disorder)
  • Lewy Body Parkinson Disease
  • PARKINSONS DIS IDIOPATHIC
  • PARKINSONS DIS
  • Parkinson's Disease, Idiopathic
  • Parkinson syndrome
  • PARKINSONS DIS LEWY BODY
  • Parkinson's disease
  • paralysis agitans
show more synonyms
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

Karyotyping (Donor)

Has the donor karyotype been analysed?
No

Donor Relations

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

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 while passaging 10 nM

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
TRA 1-60
Yes
SSEA-4
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
Passage: 57

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
No

Other Genotyping (Cell Line)

Genetic Modification#

Disease/phenotype related modifications
Parkinson's disease
Synonyms
  • Parkinson's syndrome
  • Parkinsons
  • Primary Parkinsonism
  • Parkinsons disease
  • Parkinson disease
  • Parkinson's disease (disorder)
  • Parkinson's disease NOS
  • Parkinson Disease, Idiopathic
  • PARKINSON DIS
  • Paralysis agitans
  • IDIOPATHIC PARKINSONS DIS
  • PARKINSON DIS IDIOPATHIC
  • Parkinsonism, Primary
  • Parkinson's Disease, Lewy Body
  • IDIOPATHIC PARKINSON DIS
  • Idiopathic PD
  • Idiopathic Parkinson Disease
  • Lewy Body Parkinson's Disease
  • Parkinsonian disorder
  • LEWY BODY PARKINSON DIS
  • Parkinson's
  • Idiopathic Parkinson's Disease
  • Parkinson's disease NOS (disorder)
  • Lewy Body Parkinson Disease
  • PARKINSONS DIS IDIOPATHIC
  • PARKINSONS DIS
  • Parkinson's Disease, Idiopathic
  • Parkinson syndrome
  • PARKINSONS DIS LEWY BODY
  • Parkinson's disease
  • paralysis agitans
show more synonyms
Genetic modifications
SNCA (target)
Gene knock-out
4q22.1
The SCNA triplication has been gene edited to become *putatively* normal. CRISPR/Cas was employed to create a mutation in Exon2, disrupting the ATG start site and some sequence 5' to the coding start of SCNA. PCR amplicons of the CRISPR site were cloned using the TOPO cloning technique. Sequencing of 8 TOPO clones detected 2 deletions, both of which disrupt the first ATG start site. This indicates 2 alleles were putatively knocked out, and 2 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