https://www.alphaknockout.com
Mouse Gprin2 Knockout Project (CRISPR/Cas9)
Objective: To create a Gprin2 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Gprin2 gene (NCBI Reference Sequence: NM_183209 ; Ensembl: ENSMUSG00000071531 ) is located on Mouse chromosome 14. 2 exons are identified, with the ATG start codon in exon 2 and the TGA stop codon in exon 2 (Transcript: ENSMUST00000226613). Exon 2 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:
Exon 2 starts from about 0.07% of the coding region. Exon 2 covers 100.0% of the coding region. The size of effective KO region: ~1363 bp. The KO region does not have any other known gene.
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Overview of the Targeting Strategy
Wildtype allele gRNA region 5' gRNA region 3'
1 2
Legends Exon of mouse Gprin2 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. No significant tandem repeat is found in the dot plot matrix. So this region is suitable for PCR screening or sequencing analysis.
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(21.55% 431) | C(26.25% 525) | T(26.75% 535) | G(25.45% 509)
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(21.9% 438) | C(23.45% 469) | T(27.0% 540) | G(27.65% 553)
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% chr14 - 34195812 34197811 2000 browser details YourSeq 27 509 537 2000 89.3% chr3 - 11584796 11584823 28 browser details YourSeq 27 509 537 2000 89.3% chrX + 18486660 18486687 28 browser details YourSeq 26 734 767 2000 93.4% chr12 + 70174131 70174166 36 browser details YourSeq 25 754 781 2000 96.3% chr1 - 18531512 18531541 30 browser details YourSeq 24 906 935 2000 96.2% chr1 + 89148163 89148194 32 browser details YourSeq 22 890 911 2000 100.0% chr8 - 32386901 32386922 22 browser details YourSeq 22 1191 1215 2000 96.0% chrX + 84844381 84844406 26 browser details YourSeq 22 1894 1916 2000 100.0% chr1 + 94789356 94789384 29 browser details YourSeq 20 12 31 2000 100.0% chr1 - 79561222 79561241 20
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% chr14 - 34192447 34194446 2000 browser details YourSeq 37 1462 1521 2000 95.2% chr2 + 118918050 118918396 347 browser details YourSeq 24 862 885 2000 100.0% chr1 - 115551801 115551824 24 browser details YourSeq 22 866 887 2000 100.0% chr17 + 45765830 45765851 22 browser details YourSeq 21 1857 1877 2000 100.0% chr1 - 54075092 54075112 21 browser details YourSeq 21 264 284 2000 100.0% chr3 + 109830516 109830536 21
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: Gprin2 G protein regulated inducer of neurite outgrowth 2 [ Mus musculus (house mouse) ] Gene ID: 432839, updated on 12-Aug-2019
Gene summary
Official Symbol Gprin2 provided by MGI Official Full Name G protein regulated inducer of neurite outgrowth 2 provided by MGI Primary source MGI:MGI:2444560 See related Ensembl:ENSMUSG00000071531 Gene type protein coding RefSeq status PROVISIONAL Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as Gm286; mKIAA0514; C230073P13; C130040D06Rik Expression Biased expression in CNS E18 (RPKM 2.6), whole brain E14.5 (RPKM 2.3) and 11 other tissues See more Orthologs human all
Genomic context
Location: 14; 14 B See Gprin2 in Genome Data Viewer Exon count: 5
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 14 NC_000080.6 (34185688..34201692, complement)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 14 NC_000080.5 (35007627..35014819, complement)
Chromosome 14 - NC_000080.6
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Transcript information: This gene has 3 transcripts
Gene: Gprin2 ENSMUSG00000071531
Description G protein regulated inducer of neurite outgrowth 2 [Source:MGI Symbol;Acc:MGI:2444560] Gene Synonyms C130040D06Rik Location Chromosome 14: 34,185,688-34,201,653 reverse strand. GRCm38:CM001007.2 About this gene This gene has 3 transcripts (splice variants), 107 orthologues, 1 paralogue and is a member of 1 Ensembl protein family. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Gprin2-203 ENSMUST00000226613.1 10354 455aa ENSMUSP00000154640.1 Protein coding CCDS49443 A0A2I3BRN2 GENCODE basic APPRIS P1
Gprin2-202 ENSMUST00000226511.1 438 132aa ENSMUSP00000154257.1 Protein coding - A0A2I3BQP8 CDS 3' incomplete
Gprin2-201 ENSMUST00000096019.3 430 19aa ENSMUSP00000093718.3 Protein coding - D3Z1D7 CDS 3' incomplete TSL:2
35.97 kb Forward strand 34.18Mb 34.19Mb 34.20Mb 34.21Mb Contigs < AC154738.2 Genes (Comprehensive set... < Gm18884-201processed pseudogene < Gprin2-203protein coding
< Gprin2-202protein coding
< Gprin2-201protein coding
Regulatory Build
34.18Mb 34.19Mb 34.20Mb 34.21Mb Reverse strand 35.97 kb
Regulation Legend CTCF Enhancer Promoter Promoter Flank Transcription Factor Binding Site
Gene Legend Protein Coding
merged Ensembl/Havana Ensembl protein coding
Non-Protein Coding
pseudogene
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Transcript: ENSMUST00000226613
< Gprin2-203protein coding
Reverse strand 15.95 kb
ENSMUSP00000154... MobiDB lite Low complexity (Seg) Pfam G protein-regulated inducer of neurite outgrowth, C-terminal PANTHER G protein-regulated inducer of neurite outgrowth
PTHR15718:SF5
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend missense variant synonymous variant
Scale bar 0 40 80 120 160 200 240 280 320 360 400 455
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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