Phenotypic Mutation 'Sherando' (pdf version)
Mutation Type missense
Coordinate43,058,481 bp (GRCm39)
Base Change A ⇒ G (forward strand)
Gene Siglecg
Gene Name sialic acid binding Ig-like lectin G
Synonym(s) A630096C01Rik, mSiglec-G
Chromosomal Location 43,057,623-43,067,773 bp (+) (GRCm39)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] SIGLECs are members of the immunoglobulin superfamily that are expressed on the cell surface. Most SIGLECs have 1 or more cytoplasmic immune receptor tyrosine-based inhibitory motifs, or ITIMs. SIGLECs are typically expressed on cells of the innate immune system, with the exception of the B-cell expressed SIGLEC6 (MIM 604405).[supplied by OMIM, Jul 2002]
PHENOTYPE: Mice homozygous for a null allele exhibit increased B-1 cell numbers, increased IgM levels and IgM-producing plasma cells, and produce more IgM autoantibodies. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_172900; MGI:2443630

Amino Acid Change Arginine changed to Glycine
Institutional SourceBeutler Lab
Gene Model predicted gene model for protein(s): [ENSMUSP00000005592]
AlphaFold Q80ZE3
SMART Domains Protein: ENSMUSP00000005592
Gene: ENSMUSG00000030468
AA Change: R123G

signal peptide 1 17 N/A INTRINSIC
IG 27 139 5.21e-2 SMART
IG_like 148 232 8.97e0 SMART
IGc2 262 325 3.38e-10 SMART
IGc2 366 427 8.26e-5 SMART
low complexity region 473 480 N/A INTRINSIC
transmembrane domain 545 564 N/A INTRINSIC
Predicted Effect possibly damaging

PolyPhen 2 Score 0.636 (Sensitivity: 0.87; Specificity: 0.91)
(Using ENSMUST00000005592)
Meta Mutation Damage Score 0.6898 question?
Is this an essential gene? Probably nonessential (E-score: 0.059) question?
Phenotypic Category Unknown
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Alleles Listed at MGI

All Mutations and Alleles(10) : Chemically induced (ENU)(3) Gene trapped(3) Targeted(3)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00528:Siglecg APN 7 43058481 missense possibly damaging 0.64
IGL00556:Siglecg APN 7 43061219 missense probably benign 0.02
IGL01806:Siglecg APN 7 43060888 splice site probably null
IGL01947:Siglecg APN 7 43058187 missense probably benign 0.43
IGL02257:Siglecg APN 7 43061328 missense probably benign 0.00
IGL02410:Siglecg APN 7 43058253 missense probably damaging 0.99
IGL02454:Siglecg APN 7 43058319 missense probably benign 0.00
Chamonix UTSW 7 43058846 missense possibly damaging 0.91
Dollywood UTSW 7 43060523 missense probably damaging 1.00
glowworm UTSW 7 43058003 missense probably benign 0.04
Montblanc UTSW 7 43060810 intron probably benign
Shenandoah UTSW 7 43058226 missense probably damaging 0.99
shenandoah2 UTSW 7 43061441 missense possibly damaging 0.82
Smokies UTSW 7 43058703 missense probably benign 0.02
IGL02988:Siglecg UTSW 7 43067476 missense probably damaging 1.00
R0134:Siglecg UTSW 7 43060595 missense probably damaging 1.00
R0225:Siglecg UTSW 7 43060595 missense probably damaging 1.00
R0480:Siglecg UTSW 7 43060550 missense probably benign 0.42
R1538:Siglecg UTSW 7 43067313 missense possibly damaging 0.53
R1681:Siglecg UTSW 7 43058365 missense probably benign 0.17
R2358:Siglecg UTSW 7 43058846 missense possibly damaging 0.91
R4428:Siglecg UTSW 7 43067350 missense possibly damaging 0.84
R4429:Siglecg UTSW 7 43067350 missense possibly damaging 0.84
R4736:Siglecg UTSW 7 43067332 missense probably benign 0.03
R4754:Siglecg UTSW 7 43061295 intron probably benign
R5017:Siglecg UTSW 7 43060810 intron probably benign
R5713:Siglecg UTSW 7 43058226 missense probably damaging 0.99
R5777:Siglecg UTSW 7 43058837 missense possibly damaging 0.80
R5892:Siglecg UTSW 7 43061628 intron probably benign
R6153:Siglecg UTSW 7 43061441 missense possibly damaging 0.82
R6154:Siglecg UTSW 7 43061441 missense possibly damaging 0.82
R6331:Siglecg UTSW 7 43058178 missense possibly damaging 0.83
R6562:Siglecg UTSW 7 43058481 missense possibly damaging 0.64
R6749:Siglecg UTSW 7 43058403 missense probably benign 0.00
R7066:Siglecg UTSW 7 43061166 missense probably benign 0.40
R7884:Siglecg UTSW 7 43058703 missense probably benign 0.02
R8275:Siglecg UTSW 7 43061892 missense probably benign
R8554:Siglecg UTSW 7 43058320 missense probably benign 0.01
R8846:Siglecg UTSW 7 43061942 missense probably benign 0.02
R8873:Siglecg UTSW 7 43067448 missense probably benign 0.00
R8887:Siglecg UTSW 7 43058008 missense probably benign 0.18
R9012:Siglecg UTSW 7 43060523 missense probably damaging 1.00
R9032:Siglecg UTSW 7 43061049 missense probably benign 0.24
R9048:Siglecg UTSW 7 43058003 missense probably benign 0.04
R9085:Siglecg UTSW 7 43061049 missense probably benign 0.24
R9313:Siglecg UTSW 7 43061856 missense probably benign 0.03
R9320:Siglecg UTSW 7 43058853 missense probably benign 0.33
R9745:Siglecg UTSW 7 43067476 missense probably damaging 0.98
RF006:Siglecg UTSW 7 43058288 nonsense probably null
Z1177:Siglecg UTSW 7 43061446 missense probably damaging 1.00
Mode of Inheritance Unknown
Local Stock
Last Updated 2019-09-04 9:31 PM by Diantha La Vine
Record Created 2019-01-17 11:52 AM by Bruce Beutler
Record Posted 2019-01-18
Phenotypic Description

Figure 1. Sherando mice exhibit increased frequencies of peripheral B1 cells. Flow cytometric analysis of peripheral blood was utilized to determine B1 cell frequency. Normalized data are shown. Abbreviations: WT, wild-type; REF, homozygous reference mice; HET, heterozygous variant mice; VAR, homozygous variant mice. Mean (μ) and standard deviation (σ) are indicated.

The Sherando phenotype was identified among N-ethyl-N-nitrosourea (ENU)-mutagenized G3 mice of the pedigree R6562, some of which showed increased frequencies of B1 cells in the peripheral blood (Figure 1).

Nature of Mutation

Figure 2. Linkage mapping of the increased B1 cell frequency using a recessive model of inheritance. Manhattan plot shows -log10 P values (Y-axis) plotted against the chromosome positions of 60 mutations (X-axis) identified in the G1 male of pedigree R6562. Normalized phenotype data are shown for single locus linkage analysis without consideration of G2 dam identity. Horizontal pink and red lines represent thresholds of P = 0.05, and the threshold for P = 0.05 after applying Bonferroni correction, respectively.

Whole exome HiSeq sequencing of the G1 grandsire identified 60 mutations. The increased B1 cell frequency phenotype was linked by continuous variable mapping to a mutation in Siglecg: an A to G transition at base pair 43,409,057 (v38) on chromosome 7, or base pair 865 in the GenBank genomic region NC_000073. The mutation in Siglecg was presumed causative as the B1 cell phenotype in Sherando mimics that of other Siglecg mutant alleles (see Shenandoah). Linkage was found with an additive model of inheritance, wherein four variant homozygotes and 20 heterozygous mice departed phenotypically from 15 homozygous reference mice with a P value of 4.569 x 10-5 (Figure 2). 

The mutation corresponds to residue 512 in the mRNA sequence NM_172900 within exon 3 of 12 total exons.

118 -F--F--R--M--E--R--G--F--E--R--F-

The mutated nucleotide is indicated in red. The mutation results in an arginine to glycine substitution at position 123 (R123G) in the Siglec-G protein, and is strongly predicted by PolyPhen-2 to be damaging (score = 0.636).

Illustration of Mutations in
Gene & Protein
Protein Prediction
Figure 6. Domain organization of Siglec-G. The Sherando mutation results in an arginine to glycine substitution at position 123 (R123G). Other mutations found in the Siglec-G protein are shown in red. Click on each mutation for more information.

Siglec-G (Siglec-10 in humans) is a member of the CD33-related Siglec (sialic acid–binding immunoglobulin‐like lectin) family of adhesion molecules. Siglecs specifically recognize sialic acids attached to the terminal regions of cell-surface glycoconjugates; Siglec-G preferentially binds α2,3-linked or α2,6-linked sialic acid (α2,3Sia or α2,6Sia).

Siglec-C is a type 1 transmembrane protein with a signal peptide, a sialic-binding V-set Ig-like domain, three C2-set Ig-like domains, a transmembrane domain, and a cytoplasmic tail with two putative immune receptor tyrosine-based inhibitory motifs (ITIMs) and a Grb-2 binding motif (Figure 3) (1).

The Sherando mutation results in an arginine to glycine substitution at position 123 (R123G) in the Siglec-G protein; amino acid 123 is within the V-type Ig domain.

For more information about Siglecg, please see the record Shenandoah.


Siglec-G/-10 is one of two Siglecs expressed by B cells (the other being CD22; see the record for well), and was originally identified as a B cell-associated adhesion protein that functions in the regulation of B cell activation [reviewed by (2)]. Siglec-G/-10 is a B1 cell inhibitory receptor that inhibits B cell receptor-associated NF-κB and calcium signaling, subsequently controlling the expansion and survival of B1 cells (1;3-5). The mechanism by which Siglec-G/-10 functions as an inhibitory receptor is unknown.

Siglecg-/- mice have increased levels of serum IgM and produce more IgM autoantibodies than wild-type mice (1;3). Over time, the Siglecg-/- mice develop B-cell lymphoproliferative disorders, including diffuse large B-cell lymphoma, follicular lymphoma, medium-to-large B-cell monomorphic lymphoma and atypical lymphoproliferations (6). Older Siglecg-/- mice also exhibited an autoimmune phenotype with increased autoantibody levels and mild glomerulonephritis as well as increased numbers of plasma cells, germinal center B cells, and activated CD4 T cells (7;8).

Siglecg-/- mice exhibited increased numbers of B-1 (B-1a and B-1b) B cells due to reduced spontaneous apoptosis and increased life spans of the cells (1;3-5). The increased B1 cell frequency phenotype observed in the Sherando mice indicates loss of Siglec-G-associated function.

Primers PCR Primer

Sequencing Primer

PCR program

1) 94°C 2:00
2) 94°C 0:30
3) 55°C 0:30
4) 72°C 1:00
5) repeat steps (2-4) 40x
6) 72°C 10:00
7) 4°C hold

The following sequence of 401 nucleotides is amplified (chromosome 7, + strand):

1   cctgagggaa aatggcttaa ccgttcccca ctttatggct actggttcaa aggcatcaga
61  aaaccatcac tttcatttcc agtggccaca aataacaaag ataaagtgtt agaatgggaa
121 gcccgagggc gcttccagct cttgggggat atctctaaaa agaactgttc cttgctaatc
181 aaagatgttc agtggggaga ctcaacaaac tatttcttcc ggatggagag aggatttgag
241 agattcagtt tcaaggagga gttcaggcta caagtggaag gtaagagttg tagacatagc
301 agggaacctt ggtctctcac tgggggaggg acagtggaag ataaactgtg atgcaaagaa
361 gactacttct gggaggagcc gctggagtca caggattggt a

Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red.

Science Writers Anne Murray
Illustrators Diantha La Vine
AuthorsXue Zhong, Jin Huk Choi, and Bruce Beutler