Table 1 Oligonucleotides used in this study

F: CACCGgttggatgggagagaagact

R: AAACagtcttctctcccatccaacc

-

Silencer-gRNA (Targeting silencer)

F: CACCGattgaaactaaaatctaacc

R: AAACggttagattttagtttcaatc

-

CAR-gRNA (Targeting CAR)

P1: caccgtcctgatggattagcagaac

P2: cccagaattaaaaactaatatttgctctcc

P3: caccggctctccattcaatccaaaa

P4: ccatagaaatcatttaatgggattggg

(P1 and P4):140 bp

(P2 and P4):95 bp

(P3 and P4):77 bp

RT-PCR (Detection of Non-coding RNAs)

Ovalbumin (J00895) and its promoter

F: CCCAAGCTTGGGTCGCG

ATCGCGAcctctgctttctcatatatctgtcc

R: CGGATATCCGGCCCCTTA

AGGGGtagagctgacatgatggcaayg

556

Cloning of the left homology arm

F: CCCAAGCTTGGGCTAGC

TAGCTAggtgcaaaagacagcaccaggac

R: CCGGATATCCGgtttgttctgaat cccctgttacttcc

526

Cloning of the right homology arm

F: CACCGaatgatttctatggcgtcaa

R: AAACttgacgccatagaaatcattc

-

NRE-gRNA (Targeting downstream of CAR^a)

F: CACCGtaaacttcagctagtggtat

R: AAACataccactagctgaagtttac

-

SDRE-gRNA (Targeting upstream of SDRE^b)

F: CACCGgctctagccatggtatacct

R: AAACaggtataccatggctagagc

-

OVA E2-gRNA (Targeting Ovalbumin exon 2)

P5: aatattatttgcactaccatcttgtct

P6: gtgcaagtaagagctaatatagagag

P7: cacccttaaagatacaacacatagcaca

WT^c (P5 and P7):1310

DEL^d (P5 and P7): ~ 370

WT (P5 and P6):1256

DEL (P5 and P6): ~ 316

Genomic PCR confirmation of promoter deletion

Ovalbumin (NM_205152)

P8: tgctgttgcctgatgaagtctc

P9: aatgcccatagccattaagacaga

179

RT-qPCR

GAPDH (NM_204305)

P10: cagaacatcatcccagcgtcc

P11: cagcagccttcactaccctc

187

RT-qPCR

Ovalbumin and Dsred2

P12: taccttctctctatattagctctta

P13: ggtgcttcacgtacaccttg

 ~ 2569

Genomic PCR confirmation of transgene integration