Incidental Mutation 'R1970:Als2'
ID 219606
Institutional Source Beutler Lab
Gene Symbol Als2
Ensembl Gene ENSMUSG00000026024
Gene Name alsin Rho guanine nucleotide exchange factor
Synonyms Als2cr6, 3222402C23Rik, Alsin, 9430073A21Rik
MMRRC Submission 039983-MU
Accession Numbers
Essential gene? Probably essential (E-score: 0.767) question?
Stock # R1970 (G1)
Quality Score 225
Status Validated
Chromosome 1
Chromosomal Location 59202085-59276390 bp(-) (GRCm39)
Type of Mutation missense
DNA Base Change (assembly) A to T at 59254328 bp (GRCm39)
Zygosity Heterozygous
Amino Acid Change Leucine to Glutamine at position 343 (L343Q)
Ref Sequence ENSEMBL: ENSMUSP00000125753 (fasta)
Gene Model predicted gene model for transcript(s): [ENSMUST00000027178] [ENSMUST00000160945] [ENSMUST00000163058]
AlphaFold Q920R0
Predicted Effect probably benign
Transcript: ENSMUST00000027178
AA Change: L343Q

PolyPhen 2 Score 0.341 (Sensitivity: 0.90; Specificity: 0.89)
SMART Domains Protein: ENSMUSP00000027178
Gene: ENSMUSG00000026024
AA Change: L343Q

DomainStartEndE-ValueType
Pfam:RCC1_2 93 122 1.1e-9 PFAM
Pfam:RCC1 109 165 9.5e-11 PFAM
Pfam:RCC1 170 216 6.6e-11 PFAM
low complexity region 268 282 N/A INTRINSIC
low complexity region 465 483 N/A INTRINSIC
Pfam:RCC1 521 568 5.4e-13 PFAM
Pfam:RCC1_2 555 584 8.3e-12 PFAM
Pfam:RCC1 571 619 3.4e-11 PFAM
Pfam:RhoGEF 688 877 6.5e-10 PFAM
PH 895 1001 2.17e0 SMART
MORN 1041 1062 1.34e-5 SMART
MORN 1064 1085 1.95e-1 SMART
MORN 1092 1113 6.68e-6 SMART
MORN 1115 1136 9.39e0 SMART
MORN 1143 1164 1.49e1 SMART
MORN 1167 1188 1.13e1 SMART
MORN 1190 1211 2.28e0 SMART
MORN 1213 1235 5.95e1 SMART
low complexity region 1470 1483 N/A INTRINSIC
Pfam:VPS9 1546 1650 8.6e-25 PFAM
Predicted Effect noncoding transcript
Transcript: ENSMUST00000159166
Predicted Effect probably benign
Transcript: ENSMUST00000160945
SMART Domains Protein: ENSMUSP00000140990
Gene: ENSMUSG00000026024

DomainStartEndE-ValueType
Pfam:RCC1 1 52 3.3e-8 PFAM
Pfam:RCC1_2 39 66 4.1e-5 PFAM
Predicted Effect probably benign
Transcript: ENSMUST00000163058
AA Change: L343Q

PolyPhen 2 Score 0.341 (Sensitivity: 0.90; Specificity: 0.89)
SMART Domains Protein: ENSMUSP00000125753
Gene: ENSMUSG00000026024
AA Change: L343Q

DomainStartEndE-ValueType
Pfam:RCC1_2 93 122 9.9e-10 PFAM
Pfam:RCC1 109 165 9.5e-12 PFAM
Pfam:RCC1 170 216 4.9e-12 PFAM
low complexity region 268 282 N/A INTRINSIC
low complexity region 465 483 N/A INTRINSIC
Pfam:RCC1 521 568 4.6e-14 PFAM
Pfam:RCC1_2 555 584 1.2e-11 PFAM
Pfam:RCC1 571 619 8.6e-11 PFAM
Pfam:RhoGEF 688 877 2.6e-10 PFAM
PH 895 1001 2.17e0 SMART
MORN 1041 1062 1.34e-5 SMART
MORN 1064 1085 1.95e-1 SMART
MORN 1092 1113 6.68e-6 SMART
MORN 1115 1136 9.39e0 SMART
MORN 1143 1164 1.49e1 SMART
MORN 1167 1188 1.13e1 SMART
MORN 1190 1211 2.28e0 SMART
MORN 1213 1235 5.95e1 SMART
low complexity region 1470 1483 N/A INTRINSIC
Pfam:VPS9 1546 1650 1e-24 PFAM
Predicted Effect noncoding transcript
Transcript: ENSMUST00000188469
Meta Mutation Damage Score 0.1580 question?
Coding Region Coverage
  • 1x: 99.1%
  • 3x: 98.5%
  • 10x: 96.8%
  • 20x: 94.2%
Validation Efficiency 97% (121/125)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] The protein encoded by this gene contains an ATS1/RCC1-like domain, a RhoGEF domain, and a vacuolar protein sorting 9 (VPS9) domain, all of which are guanine-nucleotide exchange factors that activate members of the Ras superfamily of GTPases. The protein functions as a guanine nucleotide exchange factor for the small GTPase RAB5. The protein localizes with RAB5 on early endosomal compartments, and functions as a modulator for endosomal dynamics. Mutations in this gene result in several forms of juvenile lateral sclerosis and infantile-onset ascending spastic paralysis. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Oct 2008]
PHENOTYPE: Homozygous null mutations in this gene may result in increased body weight, altered endosome trafficking, modest motor behavioral abnormalities, altered anxiety responses, impaired axonal transport, and mild neurolopathogical deficits including axonal degeneration in the corticospinal tract. [provided by MGI curators]
Allele List at MGI
Other mutations in this stock
Total: 121 list
GeneRefVarChr/LocMutationPredicted EffectZygosity
Abca17 A T 17: 24,526,549 (GRCm39) H578Q probably benign Het
Abcc12 A G 8: 87,253,910 (GRCm39) I958T probably benign Het
Abcg5 AATCATTTG AG 17: 84,981,030 (GRCm39) probably null Het
Acap2 A G 16: 30,952,345 (GRCm39) probably null Het
Adgrb1 T C 15: 74,411,726 (GRCm39) probably benign Het
Akap6 T A 12: 52,985,258 (GRCm39) V897E probably damaging Het
Arhgap5 A G 12: 52,589,376 (GRCm39) I1275M probably damaging Het
Arnt C T 3: 95,355,704 (GRCm39) S16L possibly damaging Het
Bglap3 C T 3: 88,284,300 (GRCm39) probably benign Het
Blnk T C 19: 40,928,609 (GRCm39) probably benign Het
C1qtnf4 T C 2: 90,720,003 (GRCm39) M92T probably damaging Het
Ccdc177 G A 12: 80,805,486 (GRCm39) R263C unknown Het
Ccdc83 T A 7: 89,893,362 (GRCm39) S132C probably damaging Het
Cdc7 A G 5: 107,120,940 (GRCm39) probably benign Het
Cgnl1 T A 9: 71,632,817 (GRCm39) N178I probably benign Het
Col27a1 C T 4: 63,191,354 (GRCm39) probably benign Het
Col5a1 A T 2: 27,876,766 (GRCm39) M822L unknown Het
Coro7 T C 16: 4,451,620 (GRCm39) I451V probably benign Het
Csmd3 T C 15: 48,536,927 (GRCm39) T92A probably damaging Het
Ddc A G 11: 11,765,292 (GRCm39) V460A possibly damaging Het
Ddx4 C T 13: 112,736,547 (GRCm39) V608I probably damaging Het
Ddx60 A T 8: 62,425,240 (GRCm39) H676L possibly damaging Het
Dmtf1 T C 5: 9,198,989 (GRCm39) E48G probably benign Het
Dpf3 T A 12: 83,371,809 (GRCm39) probably null Het
Dydc2 A G 14: 40,783,860 (GRCm39) C88R probably benign Het
Elovl4 A G 9: 83,662,772 (GRCm39) Y163H probably damaging Het
Enpp2 T C 15: 54,746,378 (GRCm39) D296G probably damaging Het
Fam83h G T 15: 75,878,419 (GRCm39) probably benign Het
Fbf1 T A 11: 116,042,317 (GRCm39) Q511L possibly damaging Het
Fhdc1 A T 3: 84,362,158 (GRCm39) L323Q probably damaging Het
Fmnl2 T A 2: 52,995,588 (GRCm39) V437D possibly damaging Het
Foxo3 A G 10: 42,073,258 (GRCm39) S420P probably benign Het
Foxp4 C T 17: 48,186,796 (GRCm39) R378Q unknown Het
Garem2 G T 5: 30,322,172 (GRCm39) G844* probably null Het
Glmp T C 3: 88,235,177 (GRCm39) L269S probably damaging Het
Gm9915 T C 1: 42,269,881 (GRCm39) noncoding transcript Het
Gnb4 A T 3: 32,652,290 (GRCm39) D27E probably damaging Het
Gnb5 A G 9: 75,251,932 (GRCm39) probably null Het
Gpr161 T G 1: 165,133,927 (GRCm39) V63G probably damaging Het
Gsk3a T A 7: 24,929,146 (GRCm39) probably benign Het
Hapln2 T C 3: 87,931,427 (GRCm39) probably null Het
Incenp G T 19: 9,862,851 (GRCm39) T401N unknown Het
Kalrn A G 16: 33,797,894 (GRCm39) probably null Het
Kcnq3 A G 15: 65,900,472 (GRCm39) probably null Het
Kif21b T C 1: 136,098,894 (GRCm39) V1394A probably damaging Het
Klhl23 T C 2: 69,664,030 (GRCm39) C460R probably damaging Het
L3mbtl1 G A 2: 162,801,492 (GRCm39) A291T probably damaging Het
Lcp1 T C 14: 75,437,946 (GRCm39) S119P probably damaging Het
Ldhc A G 7: 46,519,175 (GRCm39) I133V probably benign Het
Lmntd2 G A 7: 140,791,972 (GRCm39) probably benign Het
Lpl A T 8: 69,349,454 (GRCm39) K327* probably null Het
Lrp1b T C 2: 40,765,081 (GRCm39) D2801G probably damaging Het
Mppe1 C A 18: 67,362,843 (GRCm39) A131S probably benign Het
Msh3 T A 13: 92,386,328 (GRCm39) probably benign Het
Msh5 A G 17: 35,252,576 (GRCm39) I377T probably damaging Het
Myo1e T C 9: 70,276,055 (GRCm39) F757L probably benign Het
Myof A T 19: 37,934,082 (GRCm39) D955E probably damaging Het
Nckap1 C T 2: 80,348,286 (GRCm39) S889N probably benign Het
Ncor2 T C 5: 125,115,982 (GRCm39) D858G probably damaging Het
Neb A T 2: 52,153,917 (GRCm39) V2398D possibly damaging Het
Nefl C G 14: 68,324,121 (GRCm39) T453R probably benign Het
Nf1 A G 11: 79,444,787 (GRCm39) N371D probably benign Het
Nhsl3 A G 4: 129,119,810 (GRCm39) probably benign Het
Nlrp4f T C 13: 65,341,905 (GRCm39) Y580C probably damaging Het
Nme8 A C 13: 19,836,492 (GRCm39) L228R probably damaging Het
Numa1 C T 7: 101,658,529 (GRCm39) A1605V probably damaging Het
Ofd1 T C X: 165,210,210 (GRCm39) Y205C probably benign Het
Or2ag12 T A 7: 106,276,877 (GRCm39) N272I probably damaging Het
Or2d3b G A 7: 106,513,453 (GRCm39) G16D probably damaging Het
Or4k77 C T 2: 111,199,421 (GRCm39) S148L probably benign Het
Or8g55 T A 9: 39,785,234 (GRCm39) I221N probably damaging Het
Or8k3 T A 2: 86,058,596 (GRCm39) T240S probably damaging Het
Pclo A G 5: 14,763,487 (GRCm39) T3987A unknown Het
Pdgfrb G A 18: 61,199,566 (GRCm39) probably benign Het
Pdxk G A 10: 78,276,988 (GRCm39) T270I probably damaging Het
Pex5 T C 6: 124,391,364 (GRCm39) E10G probably damaging Het
Pik3c2g T A 6: 139,846,112 (GRCm39) probably null Het
Pkhd1 T A 1: 20,451,747 (GRCm39) I2183F probably damaging Het
Plppr2 T C 9: 21,852,422 (GRCm39) V102A probably damaging Het
Plxnd1 T C 6: 115,939,478 (GRCm39) T1449A probably damaging Het
Pnkd T A 1: 74,325,069 (GRCm39) probably null Het
Pom121l2 A G 13: 22,167,642 (GRCm39) I638V probably damaging Het
Prag1 A G 8: 36,596,314 (GRCm39) probably null Het
Ranbp10 A G 8: 106,513,340 (GRCm39) F191L probably damaging Het
Rapgef1 T C 2: 29,623,723 (GRCm39) L824P probably damaging Het
Rec8 T C 14: 55,861,599 (GRCm39) L418P probably damaging Het
Rimbp2 T A 5: 128,874,305 (GRCm39) N429Y probably damaging Het
Rpe65 C A 3: 159,321,307 (GRCm39) T373K probably benign Het
Rpl35rt T G 1: 156,194,017 (GRCm39) *124G probably null Het
Rrn3 T C 16: 13,606,938 (GRCm39) S151P probably damaging Het
Scn7a A G 2: 66,514,633 (GRCm39) V1047A possibly damaging Het
Scn9a G T 2: 66,345,724 (GRCm39) P1123Q probably damaging Het
Secisbp2l T C 2: 125,589,430 (GRCm39) D706G probably damaging Het
Serpina5 A T 12: 104,070,116 (GRCm39) T338S probably benign Het
Sez6 G A 11: 77,844,894 (GRCm39) probably null Het
Shisa4 C T 1: 135,300,012 (GRCm39) G157D probably damaging Het
Slc1a7 G T 4: 107,825,782 (GRCm39) D14Y probably benign Het
Slc25a11 A C 11: 70,536,999 (GRCm39) L51V probably benign Het
Slit3 A G 11: 35,521,668 (GRCm39) probably null Het
Spata13 G T 14: 60,928,912 (GRCm39) G157W probably damaging Het
Spta1 T C 1: 174,067,933 (GRCm39) V2120A possibly damaging Het
Srpra T C 9: 35,124,834 (GRCm39) probably null Het
Syt6 A G 3: 103,494,736 (GRCm39) I234V probably benign Het
Thada G T 17: 84,617,470 (GRCm39) P1349T probably damaging Het
Tmem161a C T 8: 70,629,559 (GRCm39) R58W probably damaging Het
Top3b A G 16: 16,701,383 (GRCm39) I232V probably damaging Het
Tspan1 T A 4: 116,020,826 (GRCm39) Q197L possibly damaging Het
Ttc28 A C 5: 111,383,501 (GRCm39) Y1334S probably benign Het
Ubxn6 G T 17: 56,380,077 (GRCm39) N28K possibly damaging Het
Uggt1 T C 1: 36,190,862 (GRCm39) D1366G probably damaging Het
Ugp2 A G 11: 21,278,942 (GRCm39) S415P probably damaging Het
Vcan A T 13: 89,837,157 (GRCm39) S2796T probably damaging Het
Vipr2 T C 12: 116,099,826 (GRCm39) V231A probably benign Het
Vmn1r176 T C 7: 23,534,373 (GRCm39) N260S probably benign Het
Vmn1r59 A T 7: 5,457,038 (GRCm39) Y241N probably damaging Het
Vmn2r67 A T 7: 84,801,013 (GRCm39) Y308N probably benign Het
Vmn2r75 A T 7: 85,797,470 (GRCm39) M781K probably damaging Het
Vwa3a A T 7: 120,379,394 (GRCm39) I500F probably damaging Het
Zfp609 A G 9: 65,702,559 (GRCm39) V31A probably damaging Het
Zfp689 G T 7: 127,043,959 (GRCm39) Q224K probably damaging Het
Zfp81 A T 17: 33,554,475 (GRCm39) L113H probably benign Het
Other mutations in Als2
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00772:Als2 APN 1 59,209,055 (GRCm39) nonsense probably null
IGL00924:Als2 APN 1 59,255,021 (GRCm39) missense probably benign 0.03
IGL00949:Als2 APN 1 59,254,731 (GRCm39) missense probably damaging 1.00
IGL00950:Als2 APN 1 59,254,541 (GRCm39) missense probably benign 0.01
IGL01090:Als2 APN 1 59,254,775 (GRCm39) missense possibly damaging 0.81
IGL01116:Als2 APN 1 59,225,163 (GRCm39) splice site probably benign
IGL02001:Als2 APN 1 59,219,347 (GRCm39) splice site probably benign
IGL02075:Als2 APN 1 59,246,945 (GRCm39) missense probably damaging 1.00
IGL02441:Als2 APN 1 59,254,631 (GRCm39) missense probably damaging 0.98
IGL02728:Als2 APN 1 59,235,506 (GRCm39) missense probably benign 0.00
IGL02740:Als2 APN 1 59,209,078 (GRCm39) missense probably benign 0.01
IGL02885:Als2 APN 1 59,206,650 (GRCm39) missense probably benign 0.30
IGL02896:Als2 APN 1 59,222,946 (GRCm39) missense probably benign 0.17
IGL02978:Als2 APN 1 59,254,324 (GRCm39) missense probably benign 0.32
IGL03032:Als2 APN 1 59,255,189 (GRCm39) splice site probably benign
IGL03065:Als2 APN 1 59,255,031 (GRCm39) missense probably benign
IGL03212:Als2 APN 1 59,242,085 (GRCm39) missense probably benign 0.00
IGL03226:Als2 APN 1 59,225,679 (GRCm39) missense probably benign 0.43
R0014:Als2 UTSW 1 59,250,547 (GRCm39) missense possibly damaging 0.53
R0243:Als2 UTSW 1 59,254,546 (GRCm39) missense probably benign
R0326:Als2 UTSW 1 59,219,742 (GRCm39) missense probably damaging 1.00
R0376:Als2 UTSW 1 59,254,724 (GRCm39) missense probably benign 0.00
R0605:Als2 UTSW 1 59,207,573 (GRCm39) missense probably benign 0.02
R1607:Als2 UTSW 1 59,219,306 (GRCm39) missense probably damaging 1.00
R1631:Als2 UTSW 1 59,257,226 (GRCm39) missense probably benign 0.00
R1657:Als2 UTSW 1 59,219,760 (GRCm39) missense probably damaging 1.00
R1763:Als2 UTSW 1 59,214,150 (GRCm39) missense probably benign
R1950:Als2 UTSW 1 59,224,760 (GRCm39) critical splice acceptor site probably null
R2151:Als2 UTSW 1 59,246,948 (GRCm39) missense probably damaging 1.00
R2292:Als2 UTSW 1 59,226,544 (GRCm39) missense probably damaging 1.00
R2513:Als2 UTSW 1 59,254,276 (GRCm39) missense probably benign 0.00
R2849:Als2 UTSW 1 59,245,697 (GRCm39) missense probably damaging 0.97
R2869:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2869:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2870:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2870:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2872:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2872:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R2873:Als2 UTSW 1 59,250,296 (GRCm39) missense probably damaging 1.00
R3054:Als2 UTSW 1 59,254,653 (GRCm39) missense probably damaging 1.00
R3081:Als2 UTSW 1 59,226,508 (GRCm39) missense probably damaging 1.00
R3176:Als2 UTSW 1 59,209,167 (GRCm39) missense possibly damaging 0.88
R3276:Als2 UTSW 1 59,209,167 (GRCm39) missense possibly damaging 0.88
R3801:Als2 UTSW 1 59,206,358 (GRCm39) missense probably damaging 1.00
R3803:Als2 UTSW 1 59,206,358 (GRCm39) missense probably damaging 1.00
R3808:Als2 UTSW 1 59,209,609 (GRCm39) missense probably benign 0.08
R3884:Als2 UTSW 1 59,224,727 (GRCm39) missense probably damaging 0.99
R4012:Als2 UTSW 1 59,226,575 (GRCm39) missense probably benign 0.09
R4033:Als2 UTSW 1 59,235,400 (GRCm39) missense probably benign
R4201:Als2 UTSW 1 59,219,313 (GRCm39) missense possibly damaging 0.77
R4321:Als2 UTSW 1 59,206,613 (GRCm39) splice site probably benign
R4707:Als2 UTSW 1 59,254,472 (GRCm39) missense probably benign
R4784:Als2 UTSW 1 59,254,472 (GRCm39) missense probably benign
R4785:Als2 UTSW 1 59,254,472 (GRCm39) missense probably benign
R4991:Als2 UTSW 1 59,246,927 (GRCm39) missense probably benign 0.10
R5068:Als2 UTSW 1 59,250,433 (GRCm39) missense probably benign 0.13
R5110:Als2 UTSW 1 59,224,600 (GRCm39) missense probably damaging 0.98
R5141:Als2 UTSW 1 59,209,611 (GRCm39) missense possibly damaging 0.80
R5394:Als2 UTSW 1 59,214,105 (GRCm39) missense probably benign 0.06
R5621:Als2 UTSW 1 59,231,049 (GRCm39) missense probably benign 0.33
R5685:Als2 UTSW 1 59,218,250 (GRCm39) missense possibly damaging 0.73
R5987:Als2 UTSW 1 59,245,746 (GRCm39) missense probably damaging 1.00
R6012:Als2 UTSW 1 59,224,374 (GRCm39) missense probably damaging 1.00
R6118:Als2 UTSW 1 59,242,228 (GRCm39) missense possibly damaging 0.62
R6222:Als2 UTSW 1 59,219,284 (GRCm39) missense probably benign 0.04
R6367:Als2 UTSW 1 59,238,299 (GRCm39) missense probably benign 0.04
R6394:Als2 UTSW 1 59,206,356 (GRCm39) missense probably damaging 0.99
R6866:Als2 UTSW 1 59,250,292 (GRCm39) missense probably damaging 1.00
R6965:Als2 UTSW 1 59,209,716 (GRCm39) missense possibly damaging 0.70
R7038:Als2 UTSW 1 59,206,673 (GRCm39) missense possibly damaging 0.94
R7178:Als2 UTSW 1 59,246,971 (GRCm39) missense probably damaging 0.96
R7494:Als2 UTSW 1 59,222,325 (GRCm39) splice site probably null
R7541:Als2 UTSW 1 59,206,775 (GRCm39) splice site probably null
R7601:Als2 UTSW 1 59,209,161 (GRCm39) missense probably benign 0.17
R8380:Als2 UTSW 1 59,250,467 (GRCm39) missense probably benign
R8478:Als2 UTSW 1 59,225,175 (GRCm39) missense probably damaging 0.96
R8492:Als2 UTSW 1 59,250,503 (GRCm39) missense probably damaging 0.98
R9048:Als2 UTSW 1 59,225,670 (GRCm39) missense possibly damaging 0.81
R9090:Als2 UTSW 1 59,242,189 (GRCm39) missense probably benign 0.01
R9128:Als2 UTSW 1 59,219,709 (GRCm39) missense probably benign 0.00
R9206:Als2 UTSW 1 59,224,406 (GRCm39) missense probably damaging 1.00
R9271:Als2 UTSW 1 59,242,189 (GRCm39) missense probably benign 0.01
R9430:Als2 UTSW 1 59,231,198 (GRCm39) missense probably benign 0.00
R9455:Als2 UTSW 1 59,219,296 (GRCm39) missense probably damaging 1.00
R9482:Als2 UTSW 1 59,231,109 (GRCm39) missense probably damaging 1.00
R9494:Als2 UTSW 1 59,206,664 (GRCm39) missense probably damaging 1.00
R9544:Als2 UTSW 1 59,250,468 (GRCm39) missense probably benign 0.00
R9796:Als2 UTSW 1 59,209,601 (GRCm39) missense probably benign 0.04
Predicted Primers PCR Primer
(F):5'- AACTTGCTGAACTTGACTTCAG -3'
(R):5'- AGAAAAGCATGCCAGCCCTG -3'

Sequencing Primer
(F):5'- CTGAACTTGACTTCAGCTCTTAAAGG -3'
(R):5'- CAGAGGCACTGGATGAGC -3'
Posted On 2014-08-25