Creation of RUES2 Cell Lines Carrying Targeted Modifications at the HTT Gene

The poster Creation of RUES2 Cell Lines Carrying Targeted Modifications at the HTT Gene presents Axol Bioscience’s collaboration with CHDI to develop a portfolio of human embryonic stem cell (hESC) lines with precise HTT gene edits for Huntington’s disease (HD) research.

Key highlights include:

• CRISPR-Cas9 Engineering: RUES2 hESCs were nucleofected with ribonucleoprotein complexes and donor vectors to introduce targeted CAG repeat expansions in HTT exon 1.
• Isogenic Clone Generation: Intermediate clones with landing pads were retargeted to produce lines with genotypes including HTT null, wild-type, high normal, and long repeat expansions.
• Genotypes & Product Codes:
  • HTT Null – CHDI-90004567, CHDI-90004568, CHDI-90004569
  • HTT WT – CHDI-90004286, CHDI-90004287, CHDI-90004288
  • WT-Retargeted (CAG22/CAG20) – CHDI-90004563, CHDI-90004564
  • CAG20/CAG22 – CHDI-90004529, CHDI-90004530
  • High Normal (CAG35/CAG22) – CHDI-90003875, CHDI-90003876
  • Long Repeats:
    • CAG98/CAG22 – CHDI-90004609
    • CAG76/CAG22 – CHDI-90004610
    • CAG118/CAG20 – CHDI-90004561, CHDI-90004562
    • CAG118/CAG22 – CHDI-90004531, CHDI-90004532
• Validation Assays:
  • Morphology (brightfield imaging)
  • Pluripotency (OCT4, NANOG, TRA-1-60)
  • Karyotyping (G-banding, aGH)
  • Cell line identity (STR analysis)
  • CAG sizing (Sanger sequencing)
• Applications: These lines can be differentiated into striatal neurons, astrocytes, and cortical excitatory neurons for use in 2D monoculture or co-culture models, supporting HD mechanism studies and preclinical drug validation.

This platform enables the HD research community to access physiologically relevant, genetically defined cell lines for translational research.

Featured Axol Products:

• RUES2 hESC Lines with HTT Modifications
• Custom Differentiation Services – Striatal neurons, astrocytes, cortical neurons from RUES2 background