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For each donor construct containing A212 or A212T, we introduced 3 silent shield mutations at the Cas9 cleavage site to protect the edited locus from subsequent cleavage

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The process includes: Resin selection and preparation Amino acid coupling Deprotection steps Washing and filtration between steps Cleavage and Deprotection After the peptide chain is complete, it needs to be cleaved from the resin and deprotected