IKMC Project Report - KOMP-CSD (ID: 28476)

Arhgap25 MGI:2443687 ENSMUSG00000030047 OTTMUSG00000023045
Pre-pipeline Designs Vectors ES Cells Mice
Mice - Genotype confirmed

Mice

Microinjection Status Allele Allele Type ES Cell Clone Genetic Background QC Data MI/Distribution Centre
currently unavailable Genotype confirmed Arhgap25tm2a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) EPD0043_2_G05 C57BL/6N view
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UCD
Southern Blot na TV Backbone Assay na 5' LR-PCR pass
Loss of WT Allele (LOA) qPCR na Homozygous Loss of WT Allele (LOA) SR-PCR na Neo Count (qPCR) na
LacZ SR-PCR na 5' Cassette Integrity Neo SR-PCR na
Mutant Specific SR-PCR na LoxP Confirmation na 3' LR-PCR
Genotyping Comment -

Mutagenesis Predictions

This gene has 4 wild type transcripts, of which 3 are protein-coding. Following removal of the floxed region, 3 transcripts are predicted to produce a truncated protein product of which 3 may be subject to non-sense mediated decay (NMD). The original allele for this mutation is of type 'Knockout First, Reporter-tagged insertion with conditional potential'. The table below shows the predicted structure of the gene transcripts after application of Flp and Cre (forming a 'Knockout-First, Post-Flp and Cre - Deletion, No Reporter' allele - more information on IKMC alleles can be found here). Click the 'view' button for each transcript to see the full prediction for that transcript.

Ensembl transcript id Ensembl Biotype Floxed transcript description Details
ENSMUST00000113637 protein_coding No protein product (NMD) view

Predicted translation:

MSLKLPRNWDFNLKAEASKIARSRSVMTGEQMAAFHPPTTPNPLERPIKVGWLKKQRSIVKNWQQRYFVLRAQQLYYYKD
EEDSKPQGCMYLPGSTVKEIATNPEEAGKFVFEVIPASSDQNRIGQDSYVLMASSQVEMEEWVKFLRRVAGTPSGAVFGQ
RLDETVAYEQKFGPHLVPILVEKCAEFILEHGVSEEGIFRLPGQDNLVKQLRDAFDAGERPSFDRLSRSW*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000696525 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000966020 Pleckstrin_homology (39/102 aa) CDS CDS
ENSMUSE00001074628 Pleckstrin_homology (30/102 aa) CDS CDS
ENSMUSE00001016708 Pleckstrin_homology (34/102 aa) CDS CDS
ENSMUSE00000968591 RhoGAP_dom (47/149 aa) CDS CDS
ENSMUSE00001060358 RhoGAP_dom (45/149 aa) CDS Deleted
ENSMUSE00001037086 RhoGAP_dom (25/149 aa) CDS CDS + stop codon + UTR
ENSMUSE00000194676 RhoGAP_dom (34/149 aa) CDS
ENSMUSE00001094657 CDS
ENSMUSE00001049410 CDS
ENSMUSE00000766189 CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000101197 protein_coding No protein product (NMD) view

Predicted translation:

MTGEQMAAFHPPTTPNPLERPIKVGWLKKQRSIVKNWQQRYFVLRAQQLYYYKDEEDSKPQGCMYLPGSTVKEIATNPEE
AGKFVFEVIPASSDQNRIGQDSYVLMASSQVEMEEWVKFLRRVAGTPSGAVFGQRLDETVAYEQKFGPHLVPILVEKCAE
FILEHGVSEEGIFRLPGQDNLVKQLRDAFDAGERPSFDRLSRSW*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000653325 UTR UTR
ENSMUSE00000978150 Pleckstrin_homology (39/102 aa) UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00001074628 Pleckstrin_homology (30/102 aa) CDS CDS
ENSMUSE00001016708 Pleckstrin_homology (34/102 aa) CDS CDS
ENSMUSE00000968591 RhoGAP_dom (47/149 aa) CDS CDS
ENSMUSE00001060358 RhoGAP_dom (45/149 aa) CDS Deleted
ENSMUSE00001037086 RhoGAP_dom (25/149 aa) CDS CDS + stop codon + UTR
ENSMUSE00000194676 RhoGAP_dom (34/149 aa) CDS
ENSMUSE00001094657 CDS
ENSMUSE00001049410 CDS
ENSMUSE00000443053 CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000071024 protein_coding No protein product (NMD) view

Predicted translation:

MYLPGSTVKEIATNPEEAGKFVFEVIPASSDQNRIGQDSYVLMASSQVEMEEWVKFLRRVAGTPSGAVFGQRLDETVAYE
QKFGPHLVPILVEKCAEFILEHGVSEEGIFRLPGQDNLVKQLRDAFDAGERPSFDRLSRSW*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000558559 UTR UTR
ENSMUSE00000970086 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00001016708 CDS CDS
ENSMUSE00000968591 RhoGAP_dom (47/149 aa) CDS CDS
ENSMUSE00001060358 RhoGAP_dom (45/149 aa) CDS Deleted
ENSMUSE00001037086 RhoGAP_dom (25/149 aa) CDS CDS + stop codon + UTR
ENSMUSE00000194676 RhoGAP_dom (34/149 aa) CDS
ENSMUSE00001094657 CDS
ENSMUSE00001049410 CDS
ENSMUSE00000443053 CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000145128 retained_intron No analysis available: incomplete transcript
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ES Cell Clones With Knockout First Mutation (Reporter Tagged Insertion With Conditional Potential)

ES Cell Clone Targeting Vector Allele Allele Type Parental ES Cell Line Genbank File Mouse QC Data
order EPD0043_2_G05 PGS00036_A_A09 Arhgap25tm2a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) JM8.F6 view yes view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype 0.7 - 0.61 Copy Number - Cells Thawed Correctly -
5' LR-PCR pass 5' SR-PCR - 3' SR-PCR pass
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot - Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -

Targeting Vectors

Design ID Vector Type Targeting Vector Cassette Backbone Genbank File
order 41908 Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) PG00036_A_4_A09 L1L2_gt2 L3L4_pZero_DTA_kan view
order 41908 Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) PG00036_A_3_A09 L1L2_gt2 L3L4_pZero_DTA_kan view
order 41908 Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) PG00036_A_2_A09 L1L2_gt2 L3L4_pZero_DTA_kan view
order 41908 Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) PG00036_A_1_A09 L1L2_gt2 L3L4_pZero_DTA_kan view
order 41908 Knockout First, Reporter-tagged insertion with conditional potential (Promotorless Cassette) PGS00036_A_A09 L1L2_gt2 L3L4_pZero_DTA_kan view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
41908 Knockout First, Reporter-tagged insertion with conditional potential PCS00036_A_A09