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. 2024 Dec 6;15:1503356. doi: 10.3389/fmicb.2024.1503356

Table 1.

The primers, reporter molecule and gRNAs used in this study.

Primers/gRNAs a Sequences and modifications b Length (nt) c Genes
ORF1ab-F1 5’-TGCATCGTGTTGTCTGT-3’ 17 ORF1ab
ORF1ab-F2 5’-TTACACCGCAAACCCGT-3’ 17
ORF1ab-CP1 5′- AGCACAAGTTGTAGGTATTTGTACATTCCCGTTGCCACATAGATCA −3’ 46
ORF1ab-CP2 5′- CGTCTGCGGTATGTGGAAAGATTGTGCATCAGCTGACTG −3’ 40
ORF1ab-C1 5’-AGCACAAGTTGTAGGTATTTGTACA-3’ 25
ORF1ab-C2 5’-CGTCTGCGGTATGTGGAAAG-3’ 20
ORF1ab-D1 5’-CACAAAATCCTTTAGGATTTG-3’ 21
ORF1ab-D2 5’-CTGTAGTTGTGATCAACTC-3’ 19
ORF1ab-R1 5’-AAAACCCACAGGGTCAT-3’ 17
ORF1ab-R2 5’-CTTAAAAACACAGTCTGTAC-3’ 20
ORF1ab-gRNA 5’-GUCUAGAGGACAGAAUUUUUCAACGGGUGUGCCAAUGGCCACUUUCCAGGUGGCAAAGCCCGUUGAGCUUCUCAAAUCUGAGAAGUGGCACCCGUUGCCACAUAGAUCAUC-3’ 111
N-F1 5’-ATTGAACCAGCTTGAGAGCA-3’ 20 N
N-F2 5’-GCAATTTGCGGCCAATGT-3’ 18
N-CP1 5’-TTGCCGAGGCTTCTTAGAAGCCAGGCCAACAACAACAAGGC-3’ 41
N-CP2 5’-AACACAAGCTTTCGGCAGACGTTGATTAGTTCCTGGTCCCCA-3’ 42
N-C1 5’-TTGCCGAGGCTTCTTAGAAGCC-3’ 22
N-C2 5’-AACACAAGCTTTCGGCAGACGT-3’ 22
N-D1 5’-CAGCAGCAGATTTCTTAGTGACAG-3’ 24
N-D2 5’-GGTCCAGAACAAACCCAAGG-3’ 20
N-R1 5’-TTTCATTGTATGCTTTAGTGGCA-3’ 23
N-gRNA 5’-GUCUAGAGGACAGAAUUUUUCAACGGGUGUGCCAAUGGCCACUUUCCAGGUGGCAAAGCCCGUUGAGCUUCUCAAAUCUGAGAAGUGGCACCAUUGUAUGCUUUAGUGGCA-3’ 111
Fluorescent reporter 5’-FAM-TTATTAT-BHQ1-3’ 7
Lateral flow reporter 5’-FAM-TTATTAT-Biotin-3’ 7
a

ORF1ab, open reading frame 1a/b; N, nucleoprotein gene.

b

ORF1ab-CP1 and N-R1primers were modified in linker region with PAM site (TTC and TTT marked in red).

c

nt, nucleitide.