ICGi022-A-1

General

Cell Line
hPSCreg name	ICGi022-A-1
Cite as:	ICGi022-A-1 (RRID:CVCL_D0VR)
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) Donor diseases: Healthy Subject ICGi022-A (K7-4Lf) Donor diseases: Healthy Subject WAe001-A-56 ICGi022-A-3 (K7-MYBPC3-N515del-1) Donor diseases: Healthy Subject ICGi022-A-4 (K7-MYBPC3-N515del-2) Donor diseases: Healthy Subject ICGi022-A-5 (K7-MYBPC3-N515del-3) Donor diseases: Healthy Subject ICGi022-A-8 (K7-4 T32) Donor diseases: Healthy Subject ICGi022-A-6 (K74-AsCas12a-N1-26) Donor diseases: Healthy Subject ICGi022-A-7 (K74-AsCas12aY1-17) Donor diseases: Healthy Subject ICGi022-B (K7-2Lf) Donor diseases: Healthy Subject WAe009-A-1P (IsoHD-30Q-NGN2) WAe009-A-1Q (IsoHD-65Q-NGN2) GIBHi002-A-1 (C5-PARK2-KO) WAe009-A-1R (IsoHD-81Q-NGN2) STBCi026-A-1 (SFC840-03-03 LRRK2-/-D10) STBCi026-A-2 (SFC840-03-03 LRRK2-/-C11) STBCi026-A-3 (SFC840-03-03 LRRK2 WT/R1441C H3) SIGi001-A-13 (iPSC0028 – MonoAllelic MAPT_Ex10+16T/Clone 1D01-11) BIONi037-A-2 (BIONi037-A ApoE2/2 #M10-7) BIONi037-A-3 (BIONi037-A ApoE3/4 #P10-22) BIONi037-A-4 (BIONi037-A ApoE4/4 #I10-53) BIONi037-A-1 (16423 ApoE KO)
Last update	24th June 2022
User feedback	Written by on No feedback available yet. Login to share your feedback, experiences or results with the research community.
Provider
Generator	Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences (ICG)
External Databases
BioSamples	SAMEA9333558
Cellosaurus	CVCL_D0VR
Wikidata	Q123032655
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	ICGi022-A

Donor Information

General Donor Information

Sex

female

Ethnicity

Caucasian

Phenotype and Disease related information (Donor)

Diseases

No disease was diagnosed.

Healthy Subject

The donor is not affected.

Synonyms

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

Donor Relations

Other cell lines of this donor

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	POU5F1 SOX2 KLF4 MYCL Trp53 LIN28
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

Supplement	Amount	Unit
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 Expression

Marker	Expressed	Immunostaining	RT-PCR	Flow Cytometry	Enzymatic Assay	Expression Profiles
NANOG	Yes	–	–
SOX2	Yes	–	–
POU5F1 (OCT-4)	Yes	–	–
TRA 1-60	Yes	–

EpiPluriScore

Score:

Marker	Present	Absent
mCpG
OCT4

Morphology

Morphology pictures

Other pluripotency tests used

Method documentation

G128R_AlkPhos.png

Differentiation Potency

Endoderm

Endoderm
Ont Id: UBERON_0000925

In vitro spontaneous differentiation

Marker	Expressed
KRT18	Yes

Morphology

Mesoderm

Mesoderm
Ont Id: UBERON_0000926

In vitro spontaneous differentiation

Marker	Expressed
ACTA2	Yes

Morphology

Ectoderm

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

Synonyms

Genetic modifications

SOD1 (target)

Type of modification

Isogenic modification

Chromosome location

21q22.11

Nucleotide sequence variant in HGVS format

NC_000021.9:c382G>C

Protein sequence variant in HGVS format

NP_000445.1:pGly128Arg

Is the modification homozygous or heterozygous?

Heterozygous

Free text

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

How has the target locus been modified?

Mutated

Available documents

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)

Type of modification

Isogenic modification

Chromosome location

21q22.11

Nucleotide sequence variant in HGVS format

NC_000021.9:c385_386delinsTG

Protein sequence variant in HGVS format

NP_000445.1:pLys129Ter

Is the modification homozygous or heterozygous?

Heterozygous

Free text

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.

How has the target locus been modified?

Mutated

Available documents

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

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

General

Cell Line

Provider

External Databases

General Information

Donor Information

General Donor Information

Phenotype and Disease related information (Donor)

Karyotyping (Donor)

Other Genotyping (Donor)

Donor Relations

External Databases (Donor)

Ethics

hIPSC Derivation

General

Reprogramming method

Vector free reprogramming

Other

Culture Conditions

Characterisation

Analysis of Undifferentiated Cells

Differentiation Potency

Microbiology / Virus Screening

Genotyping

Karyotyping (Cell Line)

Other Genotyping (Cell Line)

Genetic Modification

SOD1 (target)

SOD1 (target)