IKMC Project Report - KOMP-CSD (ID: 23401)

Nuak1 MGI:1925226 ENSMUSG00000020032 OTTMUSG00000022766
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
contact distributor Genotype confirmed Nuak1tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) EPD0088_6_D04 C57BL/6NTac;C57BL/6NTac;C57BL/6N view
( about ) 		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) pass
LacZ SR-PCR na 5' Cassette Integrity Neo SR-PCR na
Mutant Specific SR-PCR na LoxP Confirmation fail 3' LR-PCR
Genotyping Comment -

Mutagenesis Predictions

This gene has 2 wild type transcripts, of which 2 are protein-coding. Following removal of the floxed region, 2 transcripts are predicted to produce a truncated protein product of which 1 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
ENSMUST00000020220 protein_coding No protein product (NMD) view

Predicted translation: 

MEGAAVSAAGDGPAVETGLPGSPLEAVAGATAAPVEPRKPHGVKRHHHKHNLKHRYELQETLGKGTYGKVKRATERFSGR
VCLRTKIRL*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000574455 Prot_kinase_cat_dom (25/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (24/247 aa) UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000574454 Prot_kinase_cat_dom (41/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (41/247 aa) | LipoPS_kinase (37/114 aa) CDS Deleted
ENSMUSE00000574453 Prot_kinase_cat_dom (51/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (51/247 aa) | LipoPS_kinase (51/114 aa) CDS CDS + stop codon + UTR
ENSMUSE00000574452 Prot_kinase_cat_dom (22/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (22/247 aa) | LipoPS_kinase (22/114 aa) CDS
ENSMUSE00000438104 Prot_kinase_cat_dom (40/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (40/247 aa) | LipoPS_kinase (5/114 aa) CDS
ENSMUSE00000100003 Prot_kinase_cat_dom (45/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (45/247 aa) CDS
ENSMUSE00000506302 Prot_kinase_cat_dom (29/251 aa) | Ser-Thr/Tyr_kinase_cat_dom (26/247 aa) CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000129355 protein_coding Target region does not overlap 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 EPD0088_6_D04 PRPGS00031_A_A04 Nuak1tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.F6 view yes view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype 0.8 - 0.71 Copy Number pass 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 -
order EPD0088_6_E04 PRPGS00031_A_A04 Nuak1tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.F6 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype 0.8 - 0.71 Copy Number pass 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 -
order EPD0088_6_H03 PRPGS00031_A_A04 Nuak1tm1a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.F6 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype 0.9 - 0.81 Copy Number pass 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 -
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ES Cell Clones Without Conditional Potential

Note: Mutations of type "Without Conditional Potential" are correctly targeted clones that have lost the 3' LoxP site. These mutations cannot be converted into conditional alleles.

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 EPD0088_6_E03 PRPGS00031_A_A04 Nuak1tm1e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.F6 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen not confirmed 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 - 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 EPD0088_6_H04 PRPGS00031_A_A04 Nuak1tm1e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.F6 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen not confirmed 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 - 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 41745 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PRPGS00031_A_A04 L1L2_Bact_P L4L3_pD223_DTA_spec view
order 41745 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PG00031_Z_1_A04 L1L2_Bact_P L4L3_pD223_DTA_spec view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
41745 Knockout First, Reporter-tagged insertion with conditional potential PCS00031_A_A04