Compositions and methods for differential cas9 gene labeling and/or editing

A method, comprising:

• a) providing;
• i) a live cell comprising a first genomic locus and a second genomic locus and a catalytically active Cas9;
• ii) a first deoxyribonucleic acid (DNA) construct encoding a first catalytically inactive Cas9 (dCas) protein and a first fluorescent label protein having a first color, wherein said first dCas protein binds to a first single guide ribonucleic acid (sgRNA) that comprises a first complementary sequence to said first genomic locus;
• iii) a second DNA construct encoding a second dCas and a second fluorescent label protein having a second color, wherein said second dCas protein binds to a second sgRNA that comprises a second complementary sequence to said second genomic locus;
• b) transfecting said live cell with said first DNA construct, said second DNA construct, said first sgRNA and said second sgRNA, wherein said first dCas construct and said second dCas construct are expressed;
• c) detecting said first color in said first genomic locus or said second color in said second genomic locus;
• d) determining that said catalytically active Cas9 has edited:
• i) only said first genomic locus when only said first color is detected;
• ii) both said first genomic locus and said second genomic locus when both said first color and said second color is detected;
• or iii) only said second genomic locus with only said second color is detected.

A method, comprising:

• a) providing;
• i) a live cell comprising a genomic locus;
• ii) a first deoxyribonucleic acid (DNA) construct encoding a catalytically active Cas9 protein that binds to a full length single guide ribonucleic acid that comprises a first complementary sequence to said genomic locus;
• iii) a second DNA construct encoding a second catalytically active Cas9 protein and a fluorescent label protein having a color, wherein said second catalytically active Cas9 protein binds to a truncated sgRNA that comprises a second complementary sequence to said genomic locus;
• b) transfecting said live cell with said first DNA construct, said second DNA construct, said full length sgRNA and said truncated sgRNA, wherein said first dCas construct and said second dCas construct are expressed;
• c) detecting said color in said genomic locus;
• d) determining that said catalytically active Cas9 has edited said genomic locus based on said detected color.