https://www.alphaknockout.com
Mouse Rgs18 Knockout Project (CRISPR/Cas9)
Objective: To create a Rgs18 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Rgs18 gene (NCBI Reference Sequence: NM_022881 ; Ensembl: ENSMUSG00000026357 ) is located on Mouse chromosome 1. 5 exons are identified, with the ATG start codon in exon 1 and the TGA stop codon in exon 5 (Transcript: ENSMUST00000027603). Exon 2~3 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: Homozygotes for a null allele show reduced thermal nociception threshold, increased startle reflex, thrombocytopenia, defective megakaryopoiesis, and increased platelet aggregation. Homozygotes for a different null allele show decreased bleeding time, increased platelet aggregation, and thrombosis.
Exon 2 starts from about 17.02% of the coding region. Exon 2~3 covers 23.26% of the coding region. The size of effective KO region: ~1148 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 5
Legends Exon of mouse Rgs18 Knockout region
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Overview of the Dot Plot (up) Window size: 15 bp
Forward Reverse Complement
Sequence 12
Note: The 454 bp section upstream of Exon 2 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 Exon 3 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(454bp) | A(34.14% 155) | C(10.79% 49) | T(37.89% 172) | G(17.18% 78)
Note: The 454 bp section upstream of Exon 2 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(31.35% 627) | C(16.5% 330) | T(35.05% 701) | G(17.1% 342)
Note: The 2000 bp section downstream of Exon 3 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 454 1 454 454 100.0% chr1 - 144774669 144775122 454 browser details YourSeq 25 190 215 454 100.0% chr5 - 4075481 4075507 27 browser details YourSeq 24 179 204 454 96.2% chr1 + 176123753 176123778 26 browser details YourSeq 23 233 255 454 100.0% chr8 + 16727314 16727336 23 browser details YourSeq 22 136 157 454 100.0% chr13 - 59783840 59783861 22 browser details YourSeq 21 138 158 454 100.0% chr11 - 55612735 55612755 21 browser details YourSeq 21 140 160 454 100.0% chr11 - 14651067 14651087 21 browser details YourSeq 21 14 35 454 100.0% chr1 - 184976979 184977001 23 browser details YourSeq 20 139 158 454 100.0% chr10 - 101657854 101657873 20 browser details YourSeq 20 223 242 454 100.0% chr1 - 164767111 164767130 20 browser details YourSeq 20 302 321 454 100.0% chr1 - 158323197 158323216 20
Note: The 454 bp section upstream of Exon 2 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% chr1 - 144771521 144773520 2000 browser details YourSeq 172 441 674 2000 90.2% chr2 + 117672825 117673058 234 browser details YourSeq 171 442 658 2000 91.8% chrX - 41516037 41516256 220 browser details YourSeq 167 441 680 2000 89.0% chr4 - 94833042 94833306 265 browser details YourSeq 166 441 658 2000 90.3% chr4 + 116375971 116376193 223 browser details YourSeq 165 448 675 2000 89.1% chr13 - 63785692 63785929 238 browser details YourSeq 164 442 671 2000 90.3% chr3 - 58307660 58307901 242 browser details YourSeq 164 442 658 2000 91.1% chr2 + 155325121 155325341 221 browser details YourSeq 163 441 658 2000 89.9% chr11 - 18250771 18250990 220 browser details YourSeq 163 442 675 2000 89.0% chr3 + 158281957 158282192 236 browser details YourSeq 163 442 659 2000 87.6% chr12 + 91991306 92151172 159867 browser details YourSeq 161 442 666 2000 87.4% chr9 - 86102175 86102400 226 browser details YourSeq 160 441 674 2000 91.0% chr14 + 106496398 106496628 231 browser details YourSeq 159 447 671 2000 92.5% chr14 - 73169068 73169295 228 browser details YourSeq 159 441 681 2000 84.5% chrX + 73902079 73902294 216 browser details YourSeq 159 441 657 2000 88.4% chr7 + 102945204 102945418 215 browser details YourSeq 157 454 674 2000 89.5% chrX - 60193802 60194026 225 browser details YourSeq 157 442 674 2000 88.5% chr19 - 23860453 23860831 379 browser details YourSeq 157 447 658 2000 89.9% chr12 - 113965204 113965420 217 browser details YourSeq 157 441 674 2000 85.8% chr2 + 39801068 39801299 232
Note: The 2000 bp section downstream of Exon 3 is BLAT searched against the genome. No significant similarity is found.
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Gene and protein information: Rgs18 regulator of G-protein signaling 18 [ Mus musculus (house mouse) ] Gene ID: 64214, updated on 12-Aug-2019
Gene summary
Official Symbol Rgs18 provided by MGI Official Full Name regulator of G-protein signaling 18 provided by MGI Primary source MGI:MGI:1927498 See related Ensembl:ENSMUSG00000026357 Gene type protein coding RefSeq status VALIDATED Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Expression Biased expression in liver E14 (RPKM 3.7), liver E18 (RPKM 2.8) and 7 other tissues See more Orthologs human all
Genomic context
Location: 1 F; 1 62.99 cM See Rgs18 in Genome Data Viewer
Exon count: 6
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 1 NC_000067.6 (144752683..144775427, complement)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 1 NC_000067.5 (146599971..146622551, complement)
Chromosome 1 - NC_000067.6
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Transcript information: This gene has 1 transcript
Gene: Rgs18 ENSMUSG00000026357
Description regulator of G-protein signaling 18 [Source:MGI Symbol;Acc:MGI:1927498] Location Chromosome 1: 144,752,683-144,775,435 reverse strand. GRCm38:CM000994.2 About this gene This gene has 1 transcript (splice variant), 128 orthologues, 23 paralogues, is a member of 1 Ensembl protein family and is associated with 10 phenotypes. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Rgs18-201 ENSMUST00000027603.3 2031 235aa ENSMUSP00000027603.3 Protein coding CCDS15350 Q544K2 Q99PG4 TSL:1 GENCODE basic APPRIS P1
42.75 kb Forward strand 144.75Mb 144.76Mb 144.77Mb 144.78Mb Contigs < AC102163.14
Genes (Comprehensive set... < Rgs18-201protein coding
Regulatory Build
144.75Mb 144.76Mb 144.77Mb 144.78Mb Reverse strand 42.75 kb
Regulation Legend
Open Chromatin Promoter Promoter Flank Transcription Factor Binding Site
Gene Legend Protein Coding
merged Ensembl/Havana
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Transcript: ENSMUST00000027603
< Rgs18-201protein coding
Reverse strand 22.75 kb
ENSMUSP00000027... Superfamily RGS domain superfamily
SMART RGS domain
Prints RGS domain Pfam RGS domain
PROSITE profiles RGS domain
PANTHER Regulator of G-protein signalling 18
PTHR10845 Gene3D 1.10.167.10
RGS, subdomain 1/3
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend
missense variant synonymous variant
Scale bar 0 20 40 60 80 100 120 140 160 180 200 235
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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