Below are the list of genetic modifications induced by cell lines in the registry.
Homozygous insertion of sequences encoding the tdTomato fluorescent protein and a P2A peptide at the ACTA1 locus.
Genotype: tm(ACTA1t/t:ACTA1-P2A-tdTomato)
Produced by: Gene Knock-in
Correction of heterozygous ACTA1 c.443G>A (p.Gly148Asp) variant present in the parental line (MCRIi024-A). A synonymous change was also introduced (ACTA1 p.Ser147Ser). Targeted corrected clones were identified by allele-specific PCR and changes were confirmed by Sanger Sequencing.
Genotype: Homozygous: unspecified_reference_ACTA1:c.443=(p.Gly148=)
Produced by: Isogenic modification
Homozygous insertion of sequences encoding the tdTomato fluorescent protein and a P2A peptide at the C-terminal of the ACTA1 locus.
Genotype: tm(ACTA1t/t:ACTA1-P2A-tdTomato)
Produced by: Gene Knock-in
CRISPR/Cas9 was used introduce frame-shifting compound heterozygous mutations (2 bp deletion, 1 bp insertion) into exon four of the AGK gene. Alleles were confirmed by PCR and Sanger sequencing and loss of protein expression was confirmed by Western Blot. Absence of plasmid integration was confirmed by PCR. The top five predicted off-target sites were PCR amplified and Sanger sequenced, and no off-target effects were observed.
Genotype: Heterozygous: NC_000007.14:g.141596619insT
Produced by: Gene Knock-out
Correction of heterozygous NM_001232.4(CASQ2):c.539A>G((NP_001223.2):p.(Lys180Arg)) variant present in the parental line (CIAUi003-A). Clones were characterised to confirm the absence of plasmid integration, the donor variant and off-target activity of Cas9.
Genotype: NM_001232.4:c.539A=((NP_001223.2):p.(Lys180=))
Produced by: Isogenic modification
Any catecholaminergic polymorphic ventricular tachycardia in which the cause of the disease is a mutation in the CASQ2 gene.
Genotype:
Produced by: Variant
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.
Genotype: Heterozygous: NM_000086.2:c.175G=(p.Ala59=); Heterozygous: unspecified_reference_CLN3: 1.02kb deletion including of exons 7 and 8
Produced by: Isogenic modification
Introduction of heterozygous COL1A1:c.3969_3970insT variant in exon 49 leading to a frameshift and insertion of a premature stop codon. Synonymous changes were also introduced.
Genotype: Heterozygous: unspecified_reference_COL1A1:c.3969_3970insT
Produced by: Isogenic modification
Heterozygous COL2A1 c.2965C>T (p.Arg989Cys) variant was introduced to COL2A1 exon 43 to generate a model of Spondyloepiphyseal Dysplasia. Synonymous changes were also introduced.
Genotype: Heterozygous: unspecified_reference_COL2A1:c.2965C>T(p.Arg989Cys)
Produced by: Isogenic modification
Introduction of heterozygous COL2A1 c.3397GGC>TGC (p.Gly1113Cys) variant was reported.
Genotype: Heterozygous: unspecified_reference_COL2A1:c.3337G>T(p.Gly1113Cys)
Produced by: Isogenic modification