IKMC Project Report - KOMP-CSD (ID: 80199)

Ago3 MGI:2446634 ENSMUSG00000028842 OTTMUSG00000009349
Pre-pipeline Designs Vectors ES Cells Mice
Mice - Phenotype Data Available

Mice

Microinjection Status Allele Allele Type ES Cell Clone Genetic Background QC Data MI/Distribution Centre
contact distributor Genotype confirmed Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) EPD0705_1_D02 C57BL/6NTac;C57BL/6NTac;C57BL/6N-Atm1Brd/a view
( about ) 		WTSI
Southern Blot na TV Backbone Assay pass 5' LR-PCR na
Loss of WT Allele (LOA) qPCR pass Homozygous Loss of WT Allele (LOA) SR-PCR pass Neo Count (qPCR) pass
LacZ SR-PCR pass 5' Cassette Integrity Neo SR-PCR na
Mutant Specific SR-PCR pass LoxP Confirmation pass 3' LR-PCR
Genotyping Comment -

Mutagenesis Predictions

This gene has 6 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 0 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
ENSMUST00000069097 protein_coding Residual C-terminal product view

Predicted translation: 

MVQHFKVTIFGDRRPVYDGKRSLYTANPLPVATTGVDLDVTLPGEGGKDRPFKVSVKFVSRVSWHLLHEALAGGTLPEPL
ELDKPVSTNPVHAVDVVLRHLPSMKYTPVGRSFFSAPEGYDHPLGGGREVWFGFHQSVRPAMWKMMLNIDVSATAFYKAQ
PVIQFMCEVLDIHNIDEQPRPLTDSHRVKFTKEIKGLKVEVTHCGTMRRKYRVCNVTRRPASHQTFPLQLENGQTVERTV
AQYFREKYTLQLKYPHLPCLQVGQEQKHTYLPLEVCNIVAGQRCIKKLTDNQTSTMIKATARSAPDRQEEISRLVRSANY
ETDPFVQEFQFKVRDEMAHVTGRVLPAPMLQYGGRNRTVATPSHGVWDMRGKQFHTGVEIKMWAIACFATQRQCREEILK
GFTDQLRKISKDAGMPIQGQPCFCKYAQGADSVEPMFRHLKNTYSGLQLIIVILPGKTPVYAEVKRVGDTLLGMATQCVQ
VKNVIKTSPQTLSNLCLKINVKLGGINNILVPHQRPSVFQQPVIFLGADVTHPPAGDGKKPSIAAVVGSMDAHPSRYCAT
VRVQRPRQEIIQDLASMVRELLIQFYKSTRFKPTRIIFYRDGVSEGQFRQVLYYELLAIREACISLEKDYQPGITYIVVQ
KRHHTRLFCADRTERVGRSGNIPAGTTVDTDITHPYEFDFYLCSHAGIQGTSRPSHYHVLWDDNFFTADELQLLTYQLCH
TYVRCTRSVSIPAPAYYAHLVAFRARYHLVDKEHDSAEGSHVSGQSNGRDPQALAKAVQIHQDTLRTMYFA*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000788193 UTR + start codon + CDS
ENSMUSE00001091569 CDS Deleted
ENSMUSE00000631279 CDS UTR + start codon + CDS
ENSMUSE00000631278 CDS CDS
ENSMUSE00000975218 DUF1785 (44/53 aa) CDS CDS
ENSMUSE00001003440 PAZ (28/134 aa) | DUF1785 (10/53 aa) CDS CDS
ENSMUSE00000631275 PAZ (30/134 aa) CDS CDS
ENSMUSE00000631274 PAZ (50/134 aa) CDS CDS
ENSMUSE00001006869 PAZ (28/134 aa) CDS CDS
ENSMUSE00000234863 CDS CDS
ENSMUSE00000631272 CDS CDS
ENSMUSE00000384103 Piwi (12/300 aa) CDS CDS
ENSMUSE00000977673 Piwi (54/300 aa) CDS CDS
ENSMUSE00000631271 Piwi (31/300 aa) CDS CDS
ENSMUSE00000631270 Piwi (65/300 aa) CDS CDS
ENSMUSE00000631269 Piwi (45/300 aa) CDS CDS
ENSMUSE00000631268 Piwi (34/300 aa) CDS CDS
ENSMUSE00001048424 Piwi (61/300 aa) CDS CDS
ENSMUSE00000807422 CDS + stop codon + UTR CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000132123 protein_coding Target region does not overlap transcript
ENSMUST00000127831 protein_coding Target region does not overlap transcript
ENSMUST00000155645 processed_transcript Target region does not overlap transcript
ENSMUST00000151179 processed_transcript Target region does not overlap transcript
ENSMUST00000123233 processed_transcript No analysis available: incomplete transcript
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ES Cell Clones With Knockout First Mutation (Reporter Tagged Insertion With Conditional Potential)

Genome Browsers: Ensembl (mouse) - UCSC (mouse)
Tools: Southern Blot Tool - LRPCR Genotyping Primers
ES Cell Clone Targeting Vector Allele Allele Type Parental ES Cell Line Genbank File Mouse QC Data
order EPD0705_1_D02 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 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 - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 pass 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 -
order EPD0705_1_D03 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity pass
Distribution Centre
Karyotype - Copy Number pass Cells Thawed Correctly -
5' LR-PCR fail 5' SR-PCR - 3' SR-PCR fail
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 -
order EPD0705_1_F03 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen not confirmed
Loss of WT Allele (LOA) - Vector Integrity pass
Distribution Centre
Karyotype - Copy Number pass Cells Thawed Correctly -
5' LR-PCR pass 5' SR-PCR - 3' SR-PCR -
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 -
order EPD0705_1_E01 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen not confirmed LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity pass
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 -
order EPD0705_1_H02 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity pass
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 -
order EPD0705_1_G02 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen not confirmed
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 -
order EPD0705_1_G04 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen not confirmed
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 -
order EPD0705_1_H04 HTGRS6011_A_G09 Ago3tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen pass 3' Screen not confirmed
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
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 -
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Targeting Vectors

Genome Browsers: Ensembl (mouse) - UCSC (mouse)
Tools: LRPCR Genotyping Primers
Design ID Vector Type Targeting Vector Cassette Backbone Genbank File
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_4_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_1_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_8_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_6_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_7_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_5_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_2_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGR06011_A_3_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
order 36767 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) HTGRS6011_A_G09 L1L2_Bact_P R3R4_pBR_DTA+_Bsd_amp view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
36767 Knockout First, Reporter-tagged insertion with conditional potential PCS6011_A_G09