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In this final step, glutamine synthetase first reacts with gamma-glutamyl-cysteine to form an electrophilic acylphosphate intermediate by transfer of the gamma-phosphate of ATP to gamma-glutamylcysteine, and then the amino group of glycine acts as a nucleophile to attack the acylphosphate intermediate to form glutathione (28

The knock-in efficiency was determined using the equation: Knock-in (%) = (Copy number of target)/(Copy number of reference + Copy number of target) 100

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