https://www.alphaknockout.com
Mouse Mroh8 Knockout Project (CRISPR/Cas9)
Objective: To create a Mroh8 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Mroh8 gene (NCBI Reference Sequence: NM_001039557 ; Ensembl: ENSMUSG00000074627 ) is located on Mouse chromosome 2. 25 exons are identified, with the ATG start codon in exon 1 and the TAG stop codon in exon 23 (Transcript: ENSMUST00000143663). Exon 2~4 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:
Exon 2 starts from about 9.06% of the coding region. Exon 2~4 covers 12.42% of the coding region. The size of effective KO region: ~7013 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 25
Legends Exon of mouse Mroh8 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 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 4 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(29.4% 588) | C(20.45% 409) | T(30.85% 617) | G(19.3% 386)
Note: The 2000 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(27.8% 556) | C(23.4% 468) | T(28.75% 575) | G(20.05% 401)
Note: The 2000 bp section downstream of Exon 4 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% chr2 - 157272086 157274085 2000 browser details YourSeq 127 359 568 2000 86.7% chr15 - 75850591 75850798 208 browser details YourSeq 120 366 565 2000 86.6% chr1 + 75372243 75372443 201 browser details YourSeq 108 366 563 2000 88.7% chr7 - 87855722 87855921 200 browser details YourSeq 108 13 562 2000 78.8% chr2 - 91985527 91985900 374 browser details YourSeq 106 368 563 2000 84.4% chr1 - 55169787 55170108 322 browser details YourSeq 105 403 563 2000 87.9% chr14 + 44108841 44109019 179 browser details YourSeq 103 389 552 2000 87.1% chr4 - 83304708 83304887 180 browser details YourSeq 103 213 564 2000 85.9% chr14 + 59415041 59415575 535 browser details YourSeq 101 399 560 2000 85.1% chr5 + 139232053 139232241 189 browser details YourSeq 100 364 567 2000 85.8% chr12 - 82071111 82071347 237 browser details YourSeq 99 386 563 2000 83.9% chr11 + 36159692 36159868 177 browser details YourSeq 98 420 566 2000 85.6% chr4 - 117179474 117179636 163 browser details YourSeq 98 410 547 2000 85.6% chr2 + 93235912 93236049 138 browser details YourSeq 95 145 554 2000 84.7% chr5_JH584297_random - 155371 155888 518 browser details YourSeq 93 440 566 2000 87.2% chr5 + 140479501 140479627 127 browser details YourSeq 93 440 564 2000 87.2% chr11 + 54121102 54121226 125 browser details YourSeq 92 387 554 2000 86.5% chr12 - 20193716 20193881 166 browser details YourSeq 91 385 554 2000 91.9% chr4 + 119155793 119155979 187 browser details YourSeq 90 365 563 2000 78.8% chr10 - 59266954 59267143 190
Note: The 2000 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% chr2 - 157263073 157265072 2000 browser details YourSeq 279 154 636 2000 86.8% chr18 + 12717612 12718104 493 browser details YourSeq 255 159 637 2000 81.9% chr16 - 21964295 21964763 469 browser details YourSeq 246 199 648 2000 89.8% chr12 + 69530468 69531180 713 browser details YourSeq 244 159 636 2000 82.2% chr6 - 8869116 8869589 474 browser details YourSeq 235 159 636 2000 82.5% chr10 - 31526993 31527449 457 browser details YourSeq 233 159 538 2000 87.8% chr14 - 121467185 121837248 370064 browser details YourSeq 229 186 595 2000 84.0% chr8 - 127983126 127983553 428 browser details YourSeq 228 222 632 2000 85.9% chr3 - 53510642 53511055 414 browser details YourSeq 228 184 632 2000 81.2% chr10 - 114920357 114920836 480 browser details YourSeq 228 167 634 2000 87.1% chr10 - 71468623 71469110 488 browser details YourSeq 226 159 636 2000 87.4% chr2 - 161685244 161685721 478 browser details YourSeq 213 258 641 2000 85.8% chr6 - 112669535 112669915 381 browser details YourSeq 211 199 637 2000 86.0% chr1 - 89391973 89392418 446 browser details YourSeq 205 159 638 2000 86.5% chr5 + 150887012 150887482 471 browser details YourSeq 204 259 637 2000 85.3% chr10 - 109709106 109709474 369 browser details YourSeq 203 269 638 2000 87.5% chr14 + 19835993 19836376 384 browser details YourSeq 202 167 636 2000 82.8% chr7 - 57110768 57111233 466 browser details YourSeq 202 199 636 2000 80.5% chr7 - 25860008 25860405 398 browser details YourSeq 201 202 636 2000 85.1% chr9 - 51158467 51158919 453
Note: The 2000 bp section downstream of Exon 4 is BLAT searched against the genome. No significant similarity is found.
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Gene and protein information: Mroh8 maestro heat-like repeat family member 8 [ Mus musculus (house mouse) ] Gene ID: 629499, updated on 12-Aug-2019
Gene summary
Official Symbol Mroh8 provided by MGI Official Full Name maestro heat-like repeat family member 8 provided by MGI Primary source MGI:MGI:3603828 See related Ensembl:ENSMUSG00000074627 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 Also known as 4922505G16Rik Expression Biased expression in testis adult (RPKM 33.2), ovary adult (RPKM 4.4) and 8 other tissues See more Orthologs human all
Genomic context
Location: 2; 2 H1 See Mroh8 in Genome Data Viewer Exon count: 25
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 2 NC_000068.7 (157208548..157279692, complement)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 2 NC_000068.6 (157034286..157105285, complement)
Chromosome 2 - NC_000068.7
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Transcript information: This gene has 5 transcripts
Gene: Mroh8 ENSMUSG00000074627
Description maestro heat-like repeat family member 8 [Source:MGI Symbol;Acc:MGI:3603828] Gene Synonyms 4922505G16Rik Location Chromosome 2: 157,208,550-157,279,549 reverse strand. GRCm38:CM000995.2 About this gene This gene has 5 transcripts (splice variants), 122 orthologues, 10 paralogues and is a member of 1 Ensembl protein family. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Mroh8-202 ENSMUST00000143663.1 3460 1049aa ENSMUSP00000124362.1 Protein coding CCDS38303 E9PYI4 TSL:1 GENCODE basic APPRIS P1
Mroh8-205 ENSMUST00000150035.1 1701 No protein - lncRNA - - TSL:1
Mroh8-203 ENSMUST00000146848.7 575 No protein - lncRNA - - TSL:5
Mroh8-201 ENSMUST00000141081.1 528 No protein - lncRNA - - TSL:3
Mroh8-204 ENSMUST00000147602.1 469 No protein - lncRNA - - TSL:5
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91.00 kb Forward strand 157.20Mb 157.22Mb 157.24Mb 157.26Mb 157.28Mb Genes Rpn2-202 >protein coding (Comprehensive set...
Rpn2-201 >protein coding
Rpn2-208 >lncRNA
Rpn2-206 >lncRNA
Rpn2-210 >lncRNA
Rpn2-211 >lncRNA
Rpn2-205 >lncRNA
Contigs AL669828.13 > Genes (Comprehensive set... < Rbl1-201protein coding < Mroh8-201lncRNA < Mroh8-203lncRNA
< Rbl1-204retained intron < Mroh8-204lncRNA< Mroh8-205lncRNA
< Rbl1-202retained intron
< Mroh8-202protein coding
Regulatory Build
157.20Mb 157.22Mb 157.24Mb 157.26Mb 157.28Mb Reverse strand 91.00 kb
Regulation Legend CTCF Enhancer Open Chromatin Promoter Promoter Flank Transcription Factor Binding Site
Gene Legend Protein Coding
merged Ensembl/Havana Ensembl protein coding
Non-Protein Coding
RNA gene processed transcript
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Transcript: ENSMUST00000143663
< Mroh8-202protein coding
Reverse strand 71.00 kb
ENSMUSP00000124... MobiDB lite Low complexity (Seg) Superfamily Armadillo-type fold PANTHER PTHR23120:SF18
PTHR23120 Gene3D Armadillo-like helical
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend missense variant splice region variant synonymous variant
Scale bar 0 100 200 300 400 500 600 700 800 900 1049
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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