https://www.alphaknockout.com
Mouse Adcy2 Knockout Project (CRISPR/Cas9)
Objective: To create a Adcy2 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering.
Strategy summary: The Adcy2 gene (NCBI Reference Sequence: NM_153534 ; Ensembl: ENSMUSG00000021536 ) is located on Mouse chromosome 13. 25 exons are identified, with the ATG start codon in exon 1 and the TGA stop codon in exon 25 (Transcript: ENSMUST00000022013). Exon 6~10 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 26.85% of the coding region. Exon 6~10 covers 21.58% of the coding region. The size of effective KO region: ~6944 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 8 9 10 25
Legends Exon of mouse Adcy2 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 10 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.
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Overview of the GC Content Distribution (up) Window size: 300 bp
Sequence 12
Summary: Full Length(2000bp) | A(22.2% 444) | C(26.1% 522) | T(36.55% 731) | G(15.15% 303)
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(29.85% 597) | C(20.95% 419) | T(24.6% 492) | G(24.6% 492)
Note: The 2000 bp section downstream of Exon 10 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% chr13 - 68734736 68736735 2000 browser details YourSeq 172 1257 1464 2000 95.0% chr1 - 72724842 72725043 202 browser details YourSeq 169 1280 1485 2000 95.8% chr1 - 20587937 20588225 289 browser details YourSeq 168 1339 1557 2000 96.7% chr13 + 32526029 32526291 263 browser details YourSeq 157 1193 1474 2000 93.5% chrY + 18992621 18993269 649 browser details YourSeq 156 1346 1553 2000 93.4% chr3 - 74571868 74572101 234 browser details YourSeq 126 1317 1535 2000 86.7% chr11 - 110109953 110110102 150 browser details YourSeq 84 1359 1497 2000 85.4% chr15 - 89792246 89792346 101 browser details YourSeq 81 1339 1514 2000 97.7% chr1 - 36440992 36441186 195 browser details YourSeq 76 1414 1556 2000 80.8% chr1 + 64450969 64451061 93 browser details YourSeq 69 1361 1433 2000 98.7% chr1 - 81903397 81903476 80 browser details YourSeq 66 1351 1425 2000 89.8% chr1 - 70881242 70881310 69 browser details YourSeq 66 1415 1526 2000 84.6% chr15 + 69341498 69341589 92 browser details YourSeq 61 1254 1346 2000 96.9% chr11 - 54677734 54677861 128 browser details YourSeq 54 1365 1428 2000 86.5% chr18 + 54630630 54630688 59 browser details YourSeq 51 1346 1420 2000 74.6% chrX - 129017805 129017866 62 browser details YourSeq 50 1316 1389 2000 91.6% chr1 - 15303000 15303085 86 browser details YourSeq 46 1316 1375 2000 92.5% chr10 - 92072741 92072807 67 browser details YourSeq 46 1317 1383 2000 81.3% chr1 + 75669236 75669286 51 browser details YourSeq 44 1323 1471 2000 66.7% chr9 + 13745518 13745568 51
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% chr13 - 68725792 68727791 2000 browser details YourSeq 308 694 1056 2000 95.1% chr3 - 8408327 8652278 243952 browser details YourSeq 305 694 1056 2000 93.0% chr4 + 56468747 56469127 381 browser details YourSeq 304 694 1152 2000 93.7% chr2 - 108262285 108262790 506 browser details YourSeq 302 694 1056 2000 94.5% chr10 + 19512424 19512819 396 browser details YourSeq 299 694 1057 2000 92.3% chr4 + 132105738 132106114 377 browser details YourSeq 299 696 1056 2000 94.4% chr4 + 131734135 131734515 381 browser details YourSeq 295 692 1056 2000 93.3% chr2 - 5115448 5115829 382 browser details YourSeq 294 694 1056 2000 94.1% chr19 + 25215134 25215514 381 browser details YourSeq 294 694 1056 2000 93.3% chr1 + 149304797 149305176 380 browser details YourSeq 293 582 1058 2000 89.6% chr12 - 21378997 21379396 400 browser details YourSeq 293 694 1072 2000 93.3% chr17 + 64151017 64151416 400 browser details YourSeq 292 694 1056 2000 93.8% chr5 - 124212579 124212959 381 browser details YourSeq 292 698 1056 2000 94.3% chr4 - 151409687 151410063 377 browser details YourSeq 292 693 1057 2000 93.8% chr1 + 156735107 156735489 383 browser details YourSeq 291 694 1058 2000 94.5% chrX + 152035664 152036046 383 browser details YourSeq 290 693 1056 2000 95.1% chr9 - 27719386 27719761 376 browser details YourSeq 290 694 1056 2000 93.4% chr12 - 19664367 19664743 377 browser details YourSeq 290 689 1060 2000 93.4% chr1 - 88613999 88615866 1868 browser details YourSeq 290 693 1056 2000 94.0% chr10 + 103973368 103973749 382
Note: The 2000 bp section downstream of Exon 10 is BLAT searched against the genome. No significant similarity is found.
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Gene and protein information: Adcy2 adenylate cyclase 2 [ Mus musculus (house mouse) ] Gene ID: 210044, updated on 12-Aug-2019
Gene summary
Official Symbol Adcy2 provided by MGI Official Full Name adenylate cyclase 2 provided by MGI Primary source MGI:MGI:99676 See related Ensembl:ENSMUSG00000021536 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 mKIAA1060 Expression Biased expression in frontal lobe adult (RPKM 27.4), cortex adult (RPKM 24.4) and 8 other tissues See more Orthologs human all
Genomic context
Location: 13 B3; 13 35.55 cM See Adcy2 in Genome Data Viewer Exon count: 25
Annotation release Status Assembly Chr Location
108 current GRCm38.p6 (GCF_000001635.26) 13 NC_000079.6 (68620043..68999600, complement)
Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 13 NC_000079.5 (68758920..69138419, complement)
Chromosome 13 - NC_000079.6
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Transcript information: This gene has 1 transcript
Gene: Adcy2 ENSMUSG00000021536
Description adenylate cyclase 2 [Source:MGI Symbol;Acc:MGI:99676] Location Chromosome 13: 68,620,043-68,999,541 reverse strand. GRCm38:CM001006.2 About this gene This gene has 1 transcript (splice variant), 278 orthologues, 20 paralogues and is a member of 1 Ensembl protein family. Transcripts
Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags
Adcy2-201 ENSMUST00000022013.7 4211 1095aa ENSMUSP00000022013.7 Protein coding CCDS36721 Q3V1Q3 TSL:1 GENCODE basic APPRIS P1
399.50 kb Forward strand 68.7Mb 68.8Mb 68.9Mb 69.0Mb Genes 1700001L19Rik-201 >protein coding Gm26929-201 >processed pseudogene Gm35161-201 >lncRNA (Comprehensive set...
1700001L19Rik-202 >protein coding Gm7979-201 >processed pseudogene Gm35161-202 >lncRNA
Contigs < AC239879.3 AC154389.1 > < AC154482.2 Genes (Comprehensive set... < Adcy2-201protein coding
< Gm48593-201TEC
Regulatory Build
68.7Mb 68.8Mb 68.9Mb 69.0Mb Reverse strand 399.50 kb
Regulation Legend CTCF Enhancer Open Chromatin Promoter Promoter Flank Transcription Factor Binding Site
Gene Legend Protein Coding
Ensembl protein coding merged Ensembl/Havana
Non-Protein Coding
processed transcript RNA gene pseudogene
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Transcript: ENSMUST00000022013
< Adcy2-201protein coding
Reverse strand 379.50 kb
ENSMUSP00000022... Transmembrane heli... Low complexity (Seg) Coiled-coils (Ncoils) Superfamily Nucleotide cyclase SMART Adenylyl cyclase class-3/4/guanylyl cyclase Pfam Adenylyl cyclase class-3/4/guanylyl cyclase
Adenylate cyclase, N-terminal Adenylate cyclase, conserved domain PROSITE profiles Adenylyl cyclase class-3/4/guanylyl cyclase PROSITE patterns Adenylyl cyclase class-4/guanylyl cyclase, conserved site PIRSF Adenylate cyclase PANTHER PTHR45627
PTHR45627:SF6 Gene3D Nucleotide cyclase CDD cd17472 cd07302
All sequence SNPs/i... Sequence variants (dbSNP and all other sources)
Variant Legend stop gained missense variant splice region variant synonymous variant
Scale bar 0 100 200 300 400 500 600 700 800 900 1095
We wish to acknowledge the following valuable scientific information resources: Ensembl, MGI, NCBI, UCSC.
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