Table 1.
gRNAs, sequencing primers, and siRNA
PrimerSequence 5′–3′Purpose
Kras_1F 5′-TAA​TAC​GAC​TCA​CTA​TAG​GAA​TAT​AAA​CTT​GTG​GTA​GT-3′ Generation of KRAS and NRAS gRNAs 
Kras_1R 5′-TTC​TAG​CTC​TAA​AAC​ACT​ACC​ACA​AGT​TTA​TAT​TC-3′ 
Kras_2F 5′-TAA​TAC​GAC​TCA​CTA​TAG​AAT​GAC​TGA​ATA​TAA​ACT​TG-3′ 
Kras_2R 5′-TTC​TAG​CTC​TAA​AAC​CAA​GTT​TAT​ATT​CAG​TCA​TT-3′ 
Nras_1F 5′-TAA​TAC​GAC​TCA​CTA​TAG​GAC​TGA​GTA​CAA​ACT​GGT​GG-3′ 
Nras_1R 5′-TTC​TAG​CTC​TAA​AAC​CCA​CCA​GTT​TGT​ACT​CAG​TC-3′ 
Nras_2F 5′-TAA​TAC​GAC​TCA​CTA​TAG​AAT​GAC​TGA​GTA​CAA​ACT​GG-3′ 
Nras_2R 5′-TTC​TAG​CTC​TAA​AAC​CCA​GTT​TGT​ACT​CAG​TCA​TT-3′ 
KrasutrF1 5′-GCG​TCG​ATG​GAG​GAG​TTT​G-3′ Sequencing of KRAS and NRAS clones 
KrasR2 5′-ACT​CTT​GCC​TAC​GCC​ACC​A-3′ 
NrasutrF1 5′-GGG​GAG​ATC​TTT​GGA​GAC​AGA​A-3′ 
NrasR2 5′-TGG​GTA​AAG​ATG​ATC​CGA​CAA​GT-3′ 
grb2-RNA_T1 5′-ATC​TTG​TGG​AAA​GGA​CGA​AAC​ACC​GGGTT​AGA​CGT​TCC​GGT​TCA​CGGGTT​TTA​GAG​CTA​GAA​ATA​GCA​AGT​T-3′ Generation of Grb2 gRNAs 
grb2- gRNA_T2 5′-ATC​TTG​TGG​AAA​GGA​CG AAA​CAC​CGGGCT​TAG​ACG​TTC​CGG​TTC​ACGGTT​TTA​GAG​CTA​GAA​ATA​GCA​AGT​T-3′ 
Grb-HA_f 5′-TAT​CGA​TAA​GCT​TGA​TAT​CGA​GAG​GCA​GTG​TGT​AGC​CAG-3′ Generation of Grb2-mNG donor 
Grb-HA_r 5′-CTC​CTC​CTA​AGA​CGT​TCC​GGT​TCA​CGG​G-3′ 
Grb-Neon_f 5′-CCG​GAA​CGT​CTT​AGG​AGG​AGG​AGG​ATC​AG-3′ 
Grb-Neon_r 5′-TTG​ACT​CTT​ACT​TGT​ACA​GCT​CGT​CCA​TG-3′ 
Grb-3HA_f 5′-GCT​GTA​CAA​GTA​AGA​GTC​AAG​AAG​CAA​TTA​TTT​AAA​G-3′ 
Grb-3HA_r 5′-CCA​CCG​CGG​TGG​CGG​CCG​CTT​CTC​GAA​CTC​CTG​ACC​TTG-3′ 
mCh-F3 5′-ATG​GTG​AGC​AAG​GGC​GAG-3′ Generation of RAF1-mCh donor 
mCH-R2 5′-CTT​GTA​CAG​CTC​GTC​CAT​GC-3′ 
mCh_F4 5′-CTT​GCA​CGC​TGA​CCA​CGT​CCC​CGA​GGC​TGC​CTG​TCT​TCT​TAG​GAG​GA-3′ 
5′-GGA​GGA​TCA​GCA​GCA​GCA​ATG​GTG​AGC​AAG​GGC​GAG-3′ 
mCh_R3 5′-CTG​AAG​ACA​GGT​GCA​AAG​TCA​ACT​ACT​TGT​ACA​GCT​CGT​CCA​TG-3′ 
CHC siRNA 5′-GCA​ATG​AGC​TGT​TTG​AAG​A-3′ CHC knockdown 
PrimerSequence 5′–3′Purpose
Kras_1F 5′-TAA​TAC​GAC​TCA​CTA​TAG​GAA​TAT​AAA​CTT​GTG​GTA​GT-3′ Generation of KRAS and NRAS gRNAs 
Kras_1R 5′-TTC​TAG​CTC​TAA​AAC​ACT​ACC​ACA​AGT​TTA​TAT​TC-3′ 
Kras_2F 5′-TAA​TAC​GAC​TCA​CTA​TAG​AAT​GAC​TGA​ATA​TAA​ACT​TG-3′ 
Kras_2R 5′-TTC​TAG​CTC​TAA​AAC​CAA​GTT​TAT​ATT​CAG​TCA​TT-3′ 
Nras_1F 5′-TAA​TAC​GAC​TCA​CTA​TAG​GAC​TGA​GTA​CAA​ACT​GGT​GG-3′ 
Nras_1R 5′-TTC​TAG​CTC​TAA​AAC​CCA​CCA​GTT​TGT​ACT​CAG​TC-3′ 
Nras_2F 5′-TAA​TAC​GAC​TCA​CTA​TAG​AAT​GAC​TGA​GTA​CAA​ACT​GG-3′ 
Nras_2R 5′-TTC​TAG​CTC​TAA​AAC​CCA​GTT​TGT​ACT​CAG​TCA​TT-3′ 
KrasutrF1 5′-GCG​TCG​ATG​GAG​GAG​TTT​G-3′ Sequencing of KRAS and NRAS clones 
KrasR2 5′-ACT​CTT​GCC​TAC​GCC​ACC​A-3′ 
NrasutrF1 5′-GGG​GAG​ATC​TTT​GGA​GAC​AGA​A-3′ 
NrasR2 5′-TGG​GTA​AAG​ATG​ATC​CGA​CAA​GT-3′ 
grb2-RNA_T1 5′-ATC​TTG​TGG​AAA​GGA​CGA​AAC​ACC​GGGTT​AGA​CGT​TCC​GGT​TCA​CGGGTT​TTA​GAG​CTA​GAA​ATA​GCA​AGT​T-3′ Generation of Grb2 gRNAs 
grb2- gRNA_T2 5′-ATC​TTG​TGG​AAA​GGA​CG AAA​CAC​CGGGCT​TAG​ACG​TTC​CGG​TTC​ACGGTT​TTA​GAG​CTA​GAA​ATA​GCA​AGT​T-3′ 
Grb-HA_f 5′-TAT​CGA​TAA​GCT​TGA​TAT​CGA​GAG​GCA​GTG​TGT​AGC​CAG-3′ Generation of Grb2-mNG donor 
Grb-HA_r 5′-CTC​CTC​CTA​AGA​CGT​TCC​GGT​TCA​CGG​G-3′ 
Grb-Neon_f 5′-CCG​GAA​CGT​CTT​AGG​AGG​AGG​AGG​ATC​AG-3′ 
Grb-Neon_r 5′-TTG​ACT​CTT​ACT​TGT​ACA​GCT​CGT​CCA​TG-3′ 
Grb-3HA_f 5′-GCT​GTA​CAA​GTA​AGA​GTC​AAG​AAG​CAA​TTA​TTT​AAA​G-3′ 
Grb-3HA_r 5′-CCA​CCG​CGG​TGG​CGG​CCG​CTT​CTC​GAA​CTC​CTG​ACC​TTG-3′ 
mCh-F3 5′-ATG​GTG​AGC​AAG​GGC​GAG-3′ Generation of RAF1-mCh donor 
mCH-R2 5′-CTT​GTA​CAG​CTC​GTC​CAT​GC-3′ 
mCh_F4 5′-CTT​GCA​CGC​TGA​CCA​CGT​CCC​CGA​GGC​TGC​CTG​TCT​TCT​TAG​GAG​GA-3′ 
5′-GGA​GGA​TCA​GCA​GCA​GCA​ATG​GTG​AGC​AAG​GGC​GAG-3′ 
mCh_R3 5′-CTG​AAG​ACA​GGT​GCA​AAG​TCA​ACT​ACT​TGT​ACA​GCT​CGT​CCA​TG-3′ 
CHC siRNA 5′-GCA​ATG​AGC​TGT​TTG​AAG​A-3′ CHC knockdown 

Underlined sequences correspond to target gRNA sequence.

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