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ICGi022-A-1

Registration Summary:

SOD1-G128R_K7-4L-f, SOD1-G128R

ICGi022-A-1

General

Cell Line
hPSCreg Name
ICGi022-A-1
Alternative name(s)
SOD1-G128R_K7-4L-f, SOD1-G128R
Cell line type Human induced pluripotent stem cell (hiPSC)
Similar lines
ICGi022-A-2
(SOD1-D91A, SOD1-D91A_K7-4Lf)
ICGi022-A
(K7-4Lf)
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ICGi022-A-4
(K7-MYBPC3-N515del-2)
ICGi022-A-5
(K7-MYBPC3-N515del-3)
ICGi022-A-6
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ICGi022-A-7
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BIONi037-A-3
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Last update 24th June 2022
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Provider
Generator Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences (ICG)

External Databases
BioSamples SAMEA9333558
CLO CLO_0103475
Cellosaurus CVCL_D0VR

General Information
* Is the cell line readily obtainable for third parties?
Yes
Research use: allowed
Clinical use: not allowed
Commercial use: not allowed
Subclone of

Donor Information

General Donor Information
Sex female
Ethnicity Caucasian

Phenotype and Disease related information (Donor)
Diseases No disease was diagnosed.
Disease associated phenotypes no phenotypes

Karyotyping (Donor)
Has the donor karyotype been analysed?
Unknown

Other Genotyping (Donor)
Is there genome-wide genotyping or functional data available?
Yes
Exome sequencing
https://www.ncbi.nlm.nih.gov/sra/?term=SRR11413027
No disease associated mutation found

External Databases (Donor)
BioSamples SAMN14446266

Ethics

Also have a look at the ethics information for the parental line ICGi022-A .
Is there an MTA available for the cell line? No
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? IDT crRNA and tracrRNA were used for CRISPR/Cas9 gene editing
Are you aware of any constraints on the use or distribution of the cell line from the owner or any parties identified in the query above? No

hIPSC Derivation

General
The source cell information can be found in the parental cell line ICGi022-A.
Passage number reprogrammed 1

Reprogramming method
Vector type Non-integrating
Vector Episomal
Genes
Is reprogramming vector detectable?
No
Methods used
PCR
Files and images showing reprogramming vector expressed or silenced
G128R_episome.png

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

Other
Selection criteria for clones ESC-like morphology
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions

Surface coating Gelatin
Feeder cells Mouse embryonic fibroblasts
Cellfinder Ont Id: http://www.ebi.ac.uk/efo/EFO_0004040
Passage method Enzymatically
TrypLE
O2 Concentration 20 %
CO2 Concentration 5 %
Medium Other medium:
Base medium: Knock-out DMEM
Main protein source: Knock-out serum replacement
Serum concentration: 15 %
Supplements
GlutaMAX 2 mM
NEAA 0.1 mM
Penicillin-Streptomycin 100 U/ml
rhFGF basic 10 ng/ml
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Yes
Has Rock inhibitor (Y27632) been used at cryo previously with this cell line?
No
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?
Yes

Characterisation

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
NANOG
Yes
–
–
SOX2
Yes
–
–
POU5F1 (OCT-4)
Yes
–
–
TRA 1-60
Yes
–
Score:
Marker Present Absent
mCpG
OCT4
Morphology pictures
Method documentation
G128R_AlkPhos.png
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
KRT18
Yes
Morphology
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
ACTA2
Yes
Morphology
Neuron
Ont Id: CL_0000540
In vitro spontaneous differentiation
Marker Expressed
NEFH
Yes
Morphology

Microbiology / Virus Screening
Mycoplasma Negative

Genotyping

Karyotyping (Cell Line)
Has the cell line karyotype been analysed?
Yes
46XX
SOD1-G128R-Karyotype.png
Passage number: 15
Karyotyping method: G-Banding

Other Genotyping (Cell Line)

Genetic Modification

Disease/phenotype related modifications
Amyotrophic Lateral Sclerosis
Amyotrophic Lateral Sclerosis in the OLS
Synonyms
  • Amyotrophic Lateral Sclerosis
  • ALS
  • Lou Gehrig Disease
  • Amyotrophic lateral sclerosis
show more synonyms
Genetic modifications
SOD1 (target)
Isogenic modification
21q22.11
NC_000021.9:c382G>C
NP_000445.1:pGly128Arg
Heterozygous
This mutation has been linked to ALS only once (PMID: 20192886) and has been described as highly severe with rapid progression, pain syndrome, and predominant lower motor neuron implication. It has been predicted bioinformatically that this substitution induces changes in the structure of the electrostatic loop, which binds copper ions essential for both stability and enzymatic activity of SOD1 protein
Mutated
G128R allele seq.png
Part of the SOD1 gene sequence of one of the alleles. Single nucleotide substitution, which leads to the amino acid substitution is present
Genetic modifications not related to a disease
SOD1 (target)
Isogenic modification
21q22.11
NC_000021.9:c385_386delinsTG
NP_000445.1:pLys129Ter
Heterozygous
The substitution of the two amino acids in the SOD1 gene was introduced as a result of CRISPR/Cas9-induced break repair. This substitution leads to the termination codon formation and, supposedly, translation of the truncated variant of SOD1 polypeptide.
Mutated
K129Ter allele seq.png
Part of the SOD1 gene sequence of one of the alleles. Two nucleotide substitution, which leads to the termination codon formation is present