Supporting Information
Supplementary Figure S1: Pedigrees of the adRP families. Causative mutation in the PRPF31
gene, cosegregation in available family members, results of the copy-number analysis of the
minisatellite repeat element (MSR1) adjacent to the PRPF31 promoter in available symptomatic and
asymptomatic patients, and haplotype analysis in RP-0361 and RP-1599 families, are shown. m:
mutated allele. +: wild-type allele. 3: 3-copy allele of the MSR1. 4: 4-copy allele of the MSR1. *:
Hemizygous individuals for the MSR1. In blue 3 copies and in red 4 copies of the MSR1 allele. ND:
no data. (-/-): no available data. Microsatellite markers inferred are shown in brackets.
Supplementary Figure S2: Long PCR validation of the tandem duplication in family RP-0932.
Electrophoresis results of the long PCR analysis are shown. Lambda DNA/HindIII was used as
marker in a 1% agarose gel. Marker size in bp. A band of 8.6 kb, corresponding to the wild-type
allele, is observed in patients III:1, IV:1, and control sample (WT). Patients III:1 and IV:1 have an
upper band from 9.4 to 23.1 kb corresponding to the duplicated allele.
Supplementary Figure S3: Expression of the PRPF31 gene in family RP-0932, in patients with
duplication (III:1 and IV:1) and deletion (IV:12), respectively. PRPF31 expression was decreased
in patients compared to nine controls. Mean differences between both groups were significantly
different (p-value <0.001), while mean differences between patients with duplication and patient with
deletion were not significantly different (NS). Triplicate assays were performed twice.
Supplementary Figure S4. Haplotype analysis using six intragenic SNPs and five
microsatellite markers flanking PRPF31 in RP-0006 and RP-1609 families with the p.Glu181*
causative mutation. The analysis of the haplotypes showed a common region sharing a minimum
of 1.2 Mb and a maximum of 1.4 Mb defined by the alleles located along the black bar (the marker
D19S605 was not informative). Haplotypes suggest a founder effect.
1
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Supplementary Table S1. Clinical features of 26 affected patients from 15 families
characterized with a mutation in the PRPF31 gene. Typical RP fundus: pale optic disc, narrowed
retina vessels, and pigmentary changes (bone spicules). MA: macular alteration. y: years. NB: night
blindness. VFC: visual field constriction. VA: visual acuity. BCVA: best corrected visual acuity. CF:
counting finger. OD: right eye. OS: left eye. ERG: electroretinogram. NR: non-recordable. ND: no
data available. Sco: scotopic. Pho: photopic. RA: reduced amplitude. Y: yes.
2
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Supplementary Figure S1: Pedigrees of the adRP families.
RP-0182
RP-0361
m: c.1146+2T>A
I:1
I:2
RP-0006
m: p.Ile110del
I:2
m: p.Glu181*
II:5
I:1
II:1
II:4
II:2
II:3
I:1
II:5
III:1
II:2
II:3
III:3
II:4
III:7
II:1
(229)
(118)
(146)
(ND)
(+)
(93)
(113)
D19s921
D19s572
D19s927
MSR1
PRPF31
D19s418
D19s605
III:2
III:6
(227)
(135)
(137)
(3)
(+)
(97)
(111)
229
126
127
3
+
95
115
III:1
III:2
+/m
3/4
234
131
137
4
m
93
115
III:3
III:4
III:5
II:5
IV:2
IV:3
+/+
IV:1
IV:4
D19s921
D19s572
D19s927
MSR1
PRPF31
D19s418
D19s605
+/m
3/3
234
131
137
4
m
93
115
IV:2
IV:3
227
135
137
3
+
97
111
IV:4
229
118
146
ND
+
93
113
229
126
127
(3)
+
95
115
229
126
127
(3)
+
95
115
IV:6
227
135
137
(3)
+
97
111
229
126
127
(3)
+
95
115
II:2
II:6
II:7
II:3
II:4
m/+
3/3
IV:5
229
118
146
ND
+
93
113
II:1
+/+
III:1
IV:1
I:2
229
126
137
4
m
93
115
IV:7
227
135
137
3
+
97
111
III:2
III:3
III:4
III:5
III:6
III:7
III:8
III:9
IV:8
227
135
137
3
+
97
111
234
131
137
4
m
93
115
+/m
3/3
227
135
137
(3)
+
97
111
229
126
127
(3)
+
95
115
III:10
+/+
RP-0637
m: p.Thr258Aspfs*21
RP-0368
m: p.His276Glnfs*2
I:1
I:2
I:2
I:1
II:1
II:2
II:3
II:8
II:4
II:9
m/+
4/3
II:3
II:4
II:9
+/m
-/-
II:5
II:8
+/m
3/4
+/+
II:6
II:7
III:13 III:14 III:15 III:16
III:9
III:10 III:12 III:18 III:11 III:19 III:17
+/+
m/+
4/3
m/+
4/3
m/+
4/3
+/+
+/+
m/+
III:1
III:3
III:5 III:6
III:2 III:7
+/+
4/4
IV:8
IV:9
IV:10
IV:11
+/+
+/+
+/+
+/+
IV:1
II:6
II:10
+/+
II:7
II:11
+/+
II:2
+/m
3/4
+/+
m/+
4/3
II:1
II:5
+/+
IV:2 IV:3
IV:4
III:3
m/+
4/3
IV:5
III:8
III:4
+/+
IV:6 IV:7
+/+
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III:4
III:5
III:6
III:7
III:1
+/m
+/+
+/m
3/4
4/4
III:2
III:8
IV:1
IV:2
IV:3
m/+
+/+
+/+
4/3
III:9
III:14
III:12
III:13
III:10
III:11
III:15 III:16 III:17
RP-1599
RP-0777
m: p.Cys299Arg
m: del chr19:54602946 to chr19:54635178
I:1
II:1
+/+
I:1
I:2
II:2
II:1
II:3
+/+
II:4
+/+
D19s921
D19s572
D19s927
MSR1
PRPF31
D19s418
D19s605
II:5
+/m
(227)
(133)
(127)
(3)
(+)
(89)
(113)
I:2
II:2
(227)
(125)
(127)
(3)
(+)
(93)
(113)
(227)
(118)
(146)
(ND)
(+)
(93)
(113)
II:5
II:6
II:7
II:8
II:3
II:4
(236)
(133)
(127)
(4)
(m)
(93)
(113)
+/+
3/3*
III:3
III:1
III:3
III:4
m/+
+/+
3/3*
IV:1
IV:2
IV:3
III:13
III:4
227
125
127
(3)
+
93
113
D19s921
D19s572
D19s927
MSR1
PRPF31
D19s418
D19s605
III:2
III:12
III:5
III:10
III:6
227
118
146
ND
+
93
113
227
133
127
(3)
+
89
113
227
118
146
ND
+
93
113
227
125
127
3
+
93
113
III:7
III:11
III:9
227
133
127
3
+
89
113
236
133
127
4
m
93
113
III:8
236
133
127
4
m
93
113
227
133
127
(3)
+
89
113
III:1
227
133
127
3
+
89
113
227
118
146
ND
+
93
113
III:2
236
133
127
4
m
93
113
IV:4
+/+
IV:13
IV:14
IV:10
IV:11
IV:12
IV:6
IV:5
IV:7
IV:8
IV:9
IV:1
IV:2
IV:3
IV:4
m/+
4/3
RP-0932
m1: del chr19:54602946 to chr19:54632693
m2: dup chr19:54621606 to chr19:54626745
I:1
II:10
II:11
II:8
III:13
III:12
III:14
II:9
III:15
II:2
III:16
II:3
I:2
II:4
III:2
II:1
III:8
III:9
II:5
III:3
II:6
III:10
III:4
III:11
III:5
m1/+
3/3*
II:7
III:1
III:6
m2/+
4/3
+/+
III:7
IV :12 IV :13IV :14
+/m1
3/3*
IV :3
IV :4
IV :7
IV :6
IV :5
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IV :8
IV :9
IV :10
IV :11
IV :1
IV :2
RP-1609
RP-1541
RP-1970
m: p.Glu181*
m: p.Gly314Argfs*10
m: c.1146+2T>C
I:1
I:1
I:2
I:1
I:2
II:4
II:3
II:1
IV:1
IV:2
III:1
IV:3
IV:4
II:1
II:2
II:3
II:2
II:2
III:3
I:2
II:1
III:3
III:2
III:1
III:2
IV:5
III:1
IV:6
III:2
IV:1
III:3
IV:2
IV:3
IV:4
IV:5
IV:6
IV:7
V:1
V:2
+/+
+/m
3/3
V:6
V:1
V:2
V:3
V:7
V:4
V:5
IV:1
IV:2
IV:3
m/+
VI:1
VI:2
+/+
VI:3
+/m
VI:4
+/+
3/3
RP-1318
3/3
m: c.322+4_322+7del
RP-2496
m: c.1074-1G>T
RP-2426
del chr19:54618828 to chr19:54619055
RP-2136
m/+
3/3
3/3
VI:5
m/+
3/3
m/+
I:1
I:1
I:2
I:2
m: c.527+1G>T
I:1
I:1
I:2
II:1
II:2
II:1
II:3
II:2
III:2
III:5
III:3
IV:3
IV:4
II:4
I:2
II:1
II:4
II:1
II:2
II:6
II:2
II:3
II:4
II:5
II:3
+/m
III:2
III:1
3/3
III:1
III:2
III:4
III:1
IV:1
IV:2
III:7
III:6
III:3
m/+
IV:1
IV:2
III:1
III:2
3/3
+/m
+/+
3/3
+/m
3/3
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IV:5
Supplementary Figure S2: Long-PCR validation of a tandem duplication in family RP-0932.
Marker
III:1
IV:1
23130
9416
6557
4361
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WT
Supplementary Figure S3: Expression of the PRPF31 gene in family RP-0932, in patients with duplication (III:1 and IV:1) and
deletion (IV:12), respectively.
NS
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RP-0006
RP-1609
I:1
PRPF31
D19s921
D19s572
D19Ss927
rs57960425
rs2303557
c.541G>T
rs1058572
rs11556769
rs76781318
rs655240
D19s418
D19s605
226
137
127
CACA
C
G
G
C
G
C
93
114
I:2
234
135
137
CACA
C
G
A
C
G
C
89
112
234
131
135
CACA
C
T
G
C
G
C
99
112
226
133
137
CACA
C
G
G
C
G
C
101
114
I:1
D19s921
D19s572
D19s927
rs57960425
rs2303557
c.541G>T
PRPF31
rs1058572
rs11556769
rs76781318
rs655240
D19s418
D19s605
II:1
PRPF31
D19s921
D19s572
D19s927
rs57960425
rs2303557
c.541G>T
rs1058572
Rs11556769
Rs76781318
Rs655240
D19s418
D19s605
226
137
127
CACA
C
G
G
C
G
C
93
114
1.4 Mb
PRPF31
Markers/
SNPs
Start (Mb)
53.3
D19s921
53.6
D19s572
53.8
D19s927
rs57960425 c. 528-78_528-75delCACA
c.528-31C>T
rs2303557
p.Glu181*
c.541G>T
c.564G>A
rs1058572
c.735C>T
rs11556769
c.855+40G>A
rs76781318
c.1148-9T>C
rs655240
55.0
D19s418
55.2
D19s605
1.2 Mb
Supplementary Figure S4. Haplotype analysis using six intragenic SNPs and five microsatellite markers flanking PRPF31 in RP0006 and RP-1609 families with the p.Glu181* causative mutation.
222
133
137
C
G
G
C
G
C
99
116
II:2
234
131
135
CACA
C
T
G
C
G
C
99
112
234
135
137
CACA
C
G
A
C
G
C
89
112
I:2
228
122
135
CACA
C
T
G
C
G
C
99
112
D19s921
226
133
137
CACA
C
G
G
C
G
C
101
114
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D19s572
D19s927
rs57960425
rs2303557
c.541G>T
PRPF31
rs1058572
rs11556769
rs76781318
rs655240
D19s418
D19s605
226
133
137
CACA
C
G
G
C
G
C
89
112
II:1
228
122
135
CACA
C
T
G
C
G
C
99
112
226
133
137
CACA
C
G
G
C
G
C
89
112
226
133
137
CACA
C
G
G
C
G
C
95
112
Supplementary Table S1. Clinical features of 26 affected patients from 15 families characterized with mutation in the PRPF31 gene.
First symptoms and course (Age)
ID
Family
PRPF31 mutation
Age at
BCVA
Ophthalmic
Patient
NB
VFC
VA loss
Examination
OD/OS
Visual Field
ERG
Fundus aspect
Additional findings
VI:5
8
9
no
44y
0.2/0.3
Tubular
NR
Typical RP
Bilateral Cataracts (42y)
RP-0006
c.541 G>T
VII:9
5
Y
Y
19y
0.5/0.5
Tubular
NR
Typical RP
RP-0182
c.1146+2T>A
IV:4
7
Y
Y
40y
0.3/0.3
Tubular
NA
Typical RP
Bilateral Cataracts (38y)
Tubular + peripheral
RP-0361
RP-0368
c.328_330del
c.828_829del
IV:1
14
14
20
27y
0.9/0.9
islands
NR
Typical RP
IV:6
14
14
20
27y
0.7/0.7
Tubular
NA
Typical RP
IV:7
Y
Y
Y
24y
0.7/0.6
Tubular
NA
Typical RP
II:1
20
20
Y
56y
NA
NA
NA
NA
III:3
Y
Y
no
39y
1/0.6
Tubular
NA
Typical RP
II:2
18
40
50
64y
CF/CF
Tubular
NR
Typical RP + MA
OS cataract (65y)
NR
Typical RP
OS Cataract (38y)
Cataract (56y)
Photophobia. Dyschromatopsia.
Tubular + peripheral
III:1
10
20
35
40y
0.7/0.7
islands
Peripheral constriction +
RP-0637
c.770dup
IV:1
NA
NA
NA
16y
0.7/1
diffuse loss
NA
Cystoid macular oedema
RP-0777
del chr19:54602946-54635178
III:1
27
32
no
37y
0.7/0.8
Tubular
NR
Typical RP
III:1
Y
Y
Y
at old age
NA
NA
NA
NA
dup chr19:54621606-54626745
IV:1
5
7
20
30y
0.2/0.2
Tubular
NA
Typical RP + MA
cataracts (25y). OS vitrectomized
III:12
Y
14
13
51y
0.125/0.125
Tubular
NA
Typical RP + MA
Bilateral cataracts (50y)
RP-0932
del chr19:54602946-54632693
IV:12
Y
17
no
28y
0.8/0.8
Tubular
NA
Typical RP
RP-1541
c.939dup
IV:2
6
20
39
38y
0.1/0.1
NA
NA
Typical RP
Cataract (21y)
III:6
Y
7
7
43y
0.1/0.05
Tubular
NA
Typical RP
Cataract (30y)
Typical RP
Dyschromatopsia. Bilateral
RP-1599
c.895T>C
IV:4
3
5
6
15y
0.8/0.6
NA
NA
RP-1609
c.541G>T
IV:2
6
22
25
23y
0.8/0.7
Tubular
Sco:NR. Pho: RA
changes
Photophobia. Dyschromatopsia
V:1
14
12
no
16y
NA
Peripheral constriction 30º
Sco:NR. Pho: RA
NA
Hypermetropia. Astigmatism
Peripheral pigmentary
Pigmentary changes +
RP-1970
c.1146+2T>C
V:2
13
no
no
14y
1.0/1.0
Peripheral constriction 30º
Sco:NR. Pho: RA
narrowed vessels
RP-2136
c.527+1G>T
II:1
Y
Y
Y
41y
0.2/0.2
Tubular
NR
Typical RP
RP-2426
del chr19:54618828-54619055
III:1
NA
NA
NA
37y
0.8/0.8
Tubular
NA
Typical RP
RP-2496
c.1074-1G>T
III:2
20
30
35
36y
0.6/0.8
Tubular
NA
Typical RP
RP-1318
c.322+4_322+7del
III:1
9
9
16
NA
NA
NA
NA
NA
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Photophobia