https://www.alphaknockout.com
Mouse Fabp2 Knockout Project (CRISPR/Cas9)
Objective: To create a Fabp2 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Fabp2 gene (NCBI Reference Sequence: NM_007980 ; Ensembl: ENSMUSG00000023057 ) is located on Mouse chromosome 3. 4 exons are identified, with the ATG start codon in exon 1 and the TAA stop codon in exon 4 (Transcript: ENSMUST00000023820). Exon 1~4 will be selected as target site. Cas9 and gRNA will be co-injected into fertilized eggs for KO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Male mutant mice exhibit increased body weight, hyperinsulinemia, increased leptin levels on a high fat diet and have elevated circulating levels of triglycerides. Female mutant mice exhibit a decreased body weight on a high fat diet compared to controls.
Exon 1~4 covers 100.0% of the coding region. The size of effective KO region: ~4644 bp. The KO region does not have any other known gene.
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Overview of the Targeting Strategy
Wildtype allele 5' gRNA region gRNA region 3'
1 2 3 4
Legends Exon of mouse Fabp2 Knockout region
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Overview of the Dot Plot (up) Window size: 15 bp
Forward Reverse Complement
Sequence 12
Note: The 2000 bp section upstream of start codon is aligned with itself to determine if there are tandem repeats. Tandem repeats are found in the dot plot matrix. The gRNA site is selected outside of these tandem repeats.
Overview of the Dot Plot (down) Window size: 15 bp
Forward Reverse Complement
Sequence 12
Note: The 2000 bp section downstream of stop codon is aligned with itself to determine if there are tandem repeats. No significant tandem repeat is found in the dot plot matrix. So this region is suitable for PCR screening or sequencing analysis.
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Overview of the GC Content Distribution (up) Window size: 300 bp
Sequence 12
Summary: Full Length(2000bp) | A(29.6% 592) | C(21.05% 421) | T(32.3% 646) | G(17.05% 341)
Note: The 2000 bp section upstream of start codon is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis.
Overview of the GC Content Distribution (down) Window size: 300 bp
Sequence 12
Summary: Full Length(2000bp) | A(21.6% 432) | C(25.95% 519) | T(33.4% 668) | G(19.05% 381)
Note: The 2000 bp section downstream of stop codon is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis.
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BLAT Search Results (up)
QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ------browser details YourSeq 2000 1 2000 2000 100.0% chr3 + 122893414 122895413 2000 browser details YourSeq 200 102 710 2000 78.1% chr4 + 124261489 124261966 478 browser details YourSeq 188 127 707 2000 78.1% chr8 + 109115519 109116006 488 browser details YourSeq 179 102 693 2000 80.0% chr9 - 73148904 73149418 515 browser details YourSeq 178 117 698 2000 78.7% chr16 - 29158774 29159312 539 browser details YourSeq 175 139 682 2000 76.1% chr6 + 51530588 51531016 429 browser details YourSeq 175 128 710 2000 83.8% chr15 + 80815578 80816175 598 browser details YourSeq 173 102 693 2000 84.5% chr1 - 180391649 180392202 554 browser details YourSeq 173 102 698 2000 84.3% chr2 + 34949412 34949929 518 browser details YourSeq 168 130 710 2000 79.4% chr8 + 113870390 113870882 493 browser details YourSeq 164 139 693 2000 77.0% chr7 + 71243483 71243923 441 browser details YourSeq 164 183 710 2000 79.0% chr2 + 170600360 170600797 438 browser details YourSeq 162 366 708 2000 88.6% chr11 + 50945695 50946201 507 browser details YourSeq 160 289 497 2000 94.4% chr16 - 74672186 74672416 231 browser details YourSeq 160 95 685 2000 79.2% chr6 + 86590070 86590559 490 browser details YourSeq 154 102 589 2000 88.5% chr5 + 134880820 134881319 500 browser details YourSeq 153 151 709 2000 72.3% chr1 + 39167933 39168374 442 browser details YourSeq 152 104 710 2000 75.2% chr8 + 83300403 83300952 550 browser details YourSeq 147 217 695 2000 80.1% chr16 - 12182279 12182728 450 browser details YourSeq 147 103 598 2000 81.8% chr13 - 36496579 36496972 394
Note: The 2000 bp section upstream of start codon is BLAT searched against the genome. No significant similarity is found.
BLAT Search Results (down)
QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ------browser details YourSeq 2000 1 2000 2000 100.0% chr3 + 122899280 122901279 2000 browser details YourSeq 978 509 1993 2000 89.4% chr12 + 98066274 98067760 1487 browser details YourSeq 967 500 2000 2000 88.0% chr2 + 65743408 65744902 1495 browser details YourSeq 962 500 1988 2000 88.2% chr7 + 119100479 119101954 1476 browser details YourSeq 957 537 2000 2000 88.2% chr14 + 50421229 50531297 110069 browser details YourSeq 906 509 1963 2000 87.2% chr6 - 109690829 109692277 1449 browser details YourSeq 905 491 1991 2000 89.1% chr19 - 35611714 35613308 1595 browser details YourSeq 890 688 2000 2000 87.5% chr8 + 40688773 40690103 1331 browser details YourSeq 878 496 2000 2000 88.4% chr8 + 4872356 4873769 1414 browser details YourSeq 876 502 2000 2000 86.1% chr3 - 16461694 16463213 1520 browser details YourSeq 868 509 1820 2000 88.9% chr14 - 16358719 16360023 1305 browser details YourSeq 853 509 1851 2000 88.0% chr4 + 110642098 110861125 219028 browser details YourSeq 839 537 2000 2000 88.4% chr12 - 39816999 39818461 1463 browser details YourSeq 818 699 2000 2000 87.7% chr4 + 22674204 22675500 1297 browser details YourSeq 808 512 1834 2000 86.8% chr9 - 23967022 23968296 1275 browser details YourSeq 806 611 1989 2000 87.2% chr9 - 10037886 10039312 1427 browser details YourSeq 805 550 2000 2000 85.3% chrX + 44504026 44505454 1429 browser details YourSeq 804 496 1792 2000 87.8% chr5 - 128199403 128200716 1314 browser details YourSeq 801 536 1986 2000 86.1% chr1 - 44982459 44983914 1456 browser details YourSeq 801 606 2000 2000 86.1% chr7 + 87160037 87161426 1390
Note: The 2000 bp section downstream of stop codon is BLAT searched against the genome. No significant similarity is found.
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Gene and protein information: Fabp2 fatty acid binding protein 2, intestinal [ Mus musculus (house mouse) ] Gene ID: 14079, updated on 13-Aug-2019
Gene summary
Official Symbol Fabp2 provided by MGI Official Full Name fatty acid binding protein 2, intestinal provided by MGI Primary source MGI:MGI:95478 See related Ensembl:ENSMUSG00000023057 Gene type protein coding RefSeq status REVIEWED Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as Fabpi; I-FABP Summary The protein encoded by this gene is part of the fatty acid binding protein family (FABP). FABPs are a family of small, highly Expression conserved, cytoplasmic proteins that bind long-chain fatty acids and other hydrophobic ligands and participate in fatty acid uptake, transport, and metabolism. This protein functions within enterocytes, possibly to sense lipids as part of energy homeostasis. In humans polymorphisms are associated with increased fat oxidation and insulin resistance. In mice deficiency of this gene alters body weight in a gender-specific manner and causes hyperinsulinemia. [provided by RefSeq, Jan 2013] Orthologs Biased expression in large intestine adult (RPKM 347.4), small intestine adult (RPKM 41.6) and 1 other tissueS ee more human all
Genomic context
Location: 3 G1; 3 53.74 cM See Fabp2 in Genome Data Viewer
Exon count: 4
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 3 NC_000069.6 (122895072..122899506)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 3 NC_000069.5 (122598310..122602424)
Chromosome 3 - NC_000069.6
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Transcript information: This gene has 1 transcript
Gene: Fabp2 ENSMUSG00000023057
Description fatty acid binding protein 2, intestinal [Source:MGI Symbol;Acc:MGI:95478] Gene Synonyms Fabpi, I-FABP Location Chromosome 3: 122,895,072-122,899,506 forward strand. GRCm38:CM000996.2 About this gene This gene has 1 transcript (splice variant), 253 orthologues, 15 paralogues, is a member of 1 Ensembl protein family and is associated with 12 phenotypes. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Fabp2-201 ENSMUST00000023820.5 965 132aa ENSMUSP00000023820.5 Protein coding CCDS17813 P55050 Q53YP5 TSL:1 GENCODE basic APPRIS P1
24.43 kb Forward strand 122.890Mb 122.895Mb 122.900Mb 122.905Mb Genes (Comprehensive set... Fabp2-201 >protein coding
Contigs < AC155158.15 Genes < 4933405D12Rik-201lncRNA (Comprehensive set...
< 4933405D12Rik-202lncRNA
Regulatory Build
122.890Mb 122.895Mb 122.900Mb 122.905Mb Reverse strand 24.43 kb
Regulation Legend CTCF Enhancer Promoter Promoter Flank
Gene Legend Protein Coding
merged Ensembl/Havana
Non-Protein Coding
RNA gene
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Transcript: ENSMUST00000023820
4.43 kb Forward strand
Fabp2-201 >protein coding
ENSMUSP00000023... Superfamily Calycin Prints Cytosolic fatty-acid binding Pfam Lipocalin/cytosolic fatty-acid binding domain PROSITE patterns Cytosolic fatty-acid binding PANTHER Fatty acid-binding protein, intestinal
Intracellular lipid binding protein Gene3D Calycin
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend missense variant synonymous variant
Scale bar 0 20 40 60 80 100 132
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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