NBS1 I171V variant underlies individual differences in chromosomal radiosensitivity within human populations



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MRE11 binding site

RAD18 binding site

RPA binding site

FHA

BRCT1

BRCT2

754 a.a.

p.I171V (c.511A>G)

b Ovarian cancer patient 18

RNF20 binding site

ATM binding site

(NBS1I171V/+)

WT allele

Presumptive DSB site

5’…ACAATATGTGCACTCATTTGTGGACGTCCAATTGTAAAGCCAGAATATTTTACTGAATTCCTGAAAGCAG…3’

I171 (c.511A)

PAM

Cas9 recognition sequence

CRISPR/Cas9-mediated editing

ssODN (150 mer) / Targeted allele

5’…ACAATATGTGCACTCATTTGTGGACGTCCAATTGTGAAGCCGGAGTACTTTACTGAATTCCTGAAAGCAG…3’

I171 (c.511A)

Sca

 ssODN (150 mer) / Targeted allele

5’…ACAATATGTGCACTCATTTGTGGACGTCCAGTTGTGAAGCCGGAGTACTTTACTGAATTCCTGAAAGCAG…3’

V171 (c.511G)

Sca

d NBS1+/+ (parent)

e NBS1+/+ clone 1

Sca

f NBS1I171V/I171V clone 1

Sca

Fig 1

NBS1+/+

NBS1I171V/I171V

NBS1-/-

(parent) clone 1 clone 1 clone 2 clone 1 clone 2

(kDa)

NBS1

95

MRE11

81

GAPDH

37

NBS1 / GAPDH

MRE11 / GAPDH

100

98

94

91 (%)

100 106

97

115

102

98 (%)










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