https://www.alphaknockout.com
Mouse Ccdc60 Knockout Project (CRISPR/Cas9)
Objective: To create a Ccdc60 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Ccdc60 gene (NCBI Reference Sequence: NM_177759 ; Ensembl: ENSMUSG00000043913 ) is located on Mouse chromosome 5. 14 exons are identified, with the ATG start codon in exon 1 and the TAG stop codon in exon 14 (Transcript: ENSMUST00000050178). Exon 6~7 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 6 starts from about 34.31% of the coding region. Exon 6~7 covers 19.02% of the coding region. The size of effective KO region: ~6090 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 6 7 14
Legends Exon of mouse Ccdc60 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 6 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 Exon 7 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(29.2% 584) | C(23.7% 474) | T(24.0% 480) | G(23.1% 462)
Note: The 2000 bp section upstream of Exon 6 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(28.55% 571) | C(24.25% 485) | T(24.45% 489) | G(22.75% 455)
Note: The 2000 bp section downstream of Exon 7 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% chr5 - 116163574 116165573 2000 browser details YourSeq 111 569 760 2000 94.4% chr10 + 79641612 79641868 257 browser details YourSeq 107 614 765 2000 93.6% chr10 + 79641244 79642097 854 browser details YourSeq 104 614 765 2000 94.1% chr13 - 103756642 103756803 162 browser details YourSeq 101 639 796 2000 93.9% chr16 - 91421057 91421359 303 browser details YourSeq 101 617 796 2000 83.5% chr12 + 106428532 106428677 146 browser details YourSeq 100 614 765 2000 93.9% chr12 + 109062907 109063112 206 browser details YourSeq 99 614 766 2000 95.5% chr14 - 31526593 31526902 310 browser details YourSeq 99 614 765 2000 93.0% chr13 - 103756694 103757011 318 browser details YourSeq 99 641 775 2000 95.3% chr1 - 78991554 78991845 292 browser details YourSeq 98 615 761 2000 86.3% chr11 - 119583186 119583314 129 browser details YourSeq 98 614 765 2000 92.2% chr10 + 8112527 8112842 316 browser details YourSeq 97 615 760 2000 93.0% chr16 - 11625332 11625489 158 browser details YourSeq 93 634 765 2000 94.3% chr13 - 103756954 103757090 137 browser details YourSeq 90 590 712 2000 97.0% chr17 - 29739162 29739424 263 browser details YourSeq 90 614 723 2000 95.1% chr19 + 25372019 25372221 203 browser details YourSeq 89 614 725 2000 94.0% chr8 - 104391337 104391461 125 browser details YourSeq 89 615 725 2000 96.0% chr13 - 103756243 103756994 752 browser details YourSeq 88 451 712 2000 97.9% chr19 - 41633139 41633563 425 browser details YourSeq 87 590 723 2000 92.4% chr15 - 86573943 86574211 269
Note: The 2000 bp section upstream of Exon 6 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% chr5 - 116155484 116157483 2000 browser details YourSeq 336 1345 1697 2000 97.8% chr5 + 116189001 116189354 354 browser details YourSeq 245 346 648 2000 90.8% chr8 - 57770801 57771124 324 browser details YourSeq 236 346 648 2000 89.2% chrX - 8044872 8045407 536 browser details YourSeq 234 346 649 2000 88.8% chr10 + 78431265 78431590 326 browser details YourSeq 231 347 655 2000 88.0% chr11 + 95933618 95933945 328 browser details YourSeq 230 348 648 2000 89.2% chr4 + 133248984 133249298 315 browser details YourSeq 229 346 648 2000 90.3% chr2 + 4950441 4950775 335 browser details YourSeq 225 347 648 2000 88.5% chr10 - 42124011 42553790 429780 browser details YourSeq 224 346 648 2000 88.9% chr18 - 57843784 57844091 308 browser details YourSeq 224 341 649 2000 90.1% chr15 + 64623432 64623756 325 browser details YourSeq 223 347 649 2000 90.0% chr19 - 29223950 29224262 313 browser details YourSeq 223 346 649 2000 87.6% chr10 - 63356498 63356813 316 browser details YourSeq 223 364 648 2000 89.7% chr1 - 130988954 130989257 304 browser details YourSeq 223 343 648 2000 90.3% chr1 - 88485483 88485810 328 browser details YourSeq 223 346 648 2000 88.4% chrX + 85571027 85571347 321 browser details YourSeq 223 347 648 2000 88.5% chr8 + 83920125 83920459 335 browser details YourSeq 221 346 648 2000 89.4% chr8 - 23202385 23202704 320 browser details YourSeq 221 346 646 2000 87.3% chr11 - 95879450 95879768 319 browser details YourSeq 221 352 648 2000 90.8% chr1 - 134452678 134453009 332
Note: The 2000 bp section downstream of Exon 7 is BLAT searched against the genome. No significant similarity is found.
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Gene and protein information: Ccdc60 coiled-coil domain containing 60 [ Mus musculus (house mouse) ] Gene ID: 269693, updated on 24-Oct-2019
Gene summary
Official Symbol Ccdc60 provided by MGI Official Full Name coiled-coil domain containing 60 provided by MGI Primary source MGI:MGI:2141043 See related Ensembl:ENSMUSG00000043913 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 AU018954; C130098D09 Expression Restricted expression toward testis adult (RPKM 123.1) See more Orthologs human all
Genomic context
Location: 5; 5 F See Ccdc60 in Genome Data Viewer Exon count: 16
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 5 NC_000071.6 (116125581..116289000, complement)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 5 NC_000071.5 (116575590..116738994, complement)
Chromosome 5 - NC_000071.6
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Transcript information: This gene has 2 transcripts
Gene: Ccdc60 ENSMUSG00000043913
Description coiled-coil domain containing 60 [Source:MGI Symbol;Acc:MGI:2141043] Location Chromosome 5: 116,124,641-116,288,985 reverse strand. GRCm38:CM000998.2 About this gene This gene has 2 transcripts (splice variants), 110 orthologues and is a member of 1 Ensembl protein family. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Ccdc60-201 ENSMUST00000050178.12 3522 545aa ENSMUSP00000049912.6 Protein coding CCDS19599 Q8C4J0 TSL:1 GENCODE basic APPRIS P1
Ccdc60-202 ENSMUST00000086483.3 2168 207aa ENSMUSP00000083671.3 Protein coding - A0A0R4J120 TSL:1 GENCODE basic
184.34 kb Forward strand 116.12Mb 116.14Mb 116.16Mb 116.18Mb 116.20Mb 116.22Mb 116.24Mb 116.26Mb 116.28Mb Genes C330018A13Rik-201 >lncRNA Gm14507-201 >lncRNA (Comprehensive set...
Gm13837-201 >lncRNA
Contigs < AC119977.18 AC158756.4 > Genes (Comprehensive set... < Ccdc60-201protein coding
< Ccdc60-202protein coding
Regulatory Build
116.12Mb 116.14Mb 116.16Mb 116.18Mb 116.20Mb 116.22Mb 116.24Mb 116.26Mb 116.28Mb Reverse strand 184.34 kb
Regulation Legend
CTCF Enhancer Open Chromatin Promoter Promoter Flank
Gene Legend Protein Coding
merged Ensembl/Havana Ensembl protein coding
Non-Protein Coding
RNA gene
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Transcript: ENSMUST00000050178
< Ccdc60-201protein coding
Reverse strand 164.34 kb
ENSMUSP00000049... MobiDB lite Low complexity (Seg) Coiled-coils (Ncoils) Pfam Protein of unknown function DUF4698 PANTHER Protein of unknown function DUF4698
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend missense variant synonymous variant
Scale bar 0 60 120 180 240 300 360 420 480 545
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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