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. 2023 Feb 1;4(1):102084. doi: 10.1016/j.xpro.2023.102084

Figure 2.

Figure 2

Design of EGFP and mCherry donor vectors

(A and B) The EGFP and mCherry donor vectors for SIN3A bi-allelic tagging. The left homology arm is the sequence preceding the SIN3A stop codon, and the right homology arm is the sequence following the SIN3A stop codon. EGFP, mCherry, neomycin, and blasticidin sequences are used for the bi-allelic tagging of SIN3A. T2A and P2A self-cleaving peptides are used for the translation of multiple proteins from the same mRNA transcript.

(C) Example of a primer for left homology arm insertion. Silent mutations are made in the sgRNA spacer to inhibit the cutting of donor vectors by the CRISPR-Cas9 system.

(D and E) Examples of primers for right homology arm insertion. A point mutation is made in the PAM sequence to disrupt the cutting of donor vectors by the CRISPR-Cas9 system.