Errors in DNA Building Blocks Cause Unwanted Variation in Precise Gene Editing

Synthesis errors are present in all single-stranded oligonucleotides (ssODNs) tested at rates that vary more than two-fold among manufacturers.

• Synthesis errors occur in ssODNs, affecting their sequence fidelity.
• The rates of errors vary significantly depending on the manufacturer and the specific sequence context.
• Errors in ssODNs are incorporated into the genome during gene editing at frequencies that reflect their initial abundance.
• In a sickle cell mutation correction protocol, common synthesis errors may lead to benign variants, while rarer errors could result in severe genetic conditions.
• Current quality control methods are inadequate for detecting these synthesis errors, indicating a need for enhanced oversight.

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