GOTI

GOTI identifies genome-wide off-target variants induced by genome editing in mouse two-cell embryos to assess the precision of CRISPR-Cas9 and base editors.

Key Features:

• High Sensitivity Detection: By comparing edited and non-edited cells from one-blastomere-injected two-cell mouse embryos, GOTI detects off-target mutations while eliminating genetic background variation.
• Versatility Across Editing Tools: Detects off-target variants induced by genome editing technologies, including CRISPR-Cas9 and base editors.
• Experimental and Computational Integration: Combines two-cell embryo injection, sampling at embryonic day 14.5, fluorescence-activated cell sorting (FACS), and whole-genome sequencing to link experimental contrast with computational analysis.
• GOTI-seq Computational Workflow: Processes raw whole-genome sequencing data and automates generation of genome-wide off-target variant reports.

Scientific Applications:

• Safety Evaluation of Genome Editing Tools: Identifies unintended mutations to assess precision and potential risks of genome editing technologies.
• Development of Gene Therapy Protocols: Detects unintended genetic modifications to inform safer gene therapy strategies.
• Basic Research in Genetic Engineering: Enables study of mechanisms and consequences of off-target effects in genome editing experiments.

Methodology:

GOTI-seq processes raw whole-genome sequencing data and generates genome-wide off-target variant reports.