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T7-NLS hCas9-pA

Cas9-poly(A) expressing improved plasmid.

Catalog number RDB13130
Resource name T7-NLS hCas9-pA
Clone info. Cas9 expressing plasmid. Induction with CMV or T7 promoter. Applicable for both direct transfection into mammalian cells and T7 in-vitro translation.
Digest with Nhe I for linearization prior to in vitro transcription followed by capping and polyadenylation.
Vector backbone pcDNA3 (Plasmid)
Selectable markers Amp^r
Gene/insert name Streptococcus pyogenes Cas9 genomic DNA
Depositor|Developer Mashimo, Tomoji |
 
Other clones in our bank Streptococcus pyogenes (KEGG orthology K09952) |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Yoshimi, K. et al. Nat. Commun. 7: 10431, 2016). RECIPIENT which wants to use the BIOLOGICAL RESOURCE for the purpose other than education or not-for-profit research is requested to enter into a Material Transfer Agreement with Osaka University.
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Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose) [Word]
Remarks
The BIOLOGICAL RESOURCE is not provided to for-profit organization or for for-profit research by non-profit organizations. Please refer the Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose).
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RDB13130 T7-NLS hCas9-pA DNA solution

Ordering Information [in Japanese] [in English]
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

References and tips

Electronic file

Sequence (full) RDB13130zzt02.seq compiled by us

Original reference

original Yoshimi, K., ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes. Nat. Commun. 7: 10431 (2016). PMID 26786405.

Further references such as user reports and related articles (go to bottom)

Featured content

Featured content Genome Editing (English text)
Featured content Genome Editing (Japanese text)

Sequence information

RIKEN BRC has sequenced portions of this material for quality test.
Please review the QC test results indicated by check icon as well as clone information before placing your order.

Test sheet RDB13130_A5Bkp1-2.pdf check

Nucleotide sequence of a portion of this resource (if available).

Primer: CMV_Forward
Region: SV40 NLS, insert 5'(D10 WT)
Sequence file: RDB13130_A5Bkd.seq check
>13130_13130_A5Bk_CMV_Foward_3_H02_23.ab1
    1 NNNNNNNNNN NNNCNNNTNG NNGCNNNTGG GCGGTAGGCG TGNNNNNNGG GAGGTCTATA
   61 TAAGCAGAGC TCGTTNAGNG AACCGTCAGA TCGCCTGGAG ACGCCATCCA CGCTGTTTTG
  121 ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGGAC TCTAGTAATA CGACTCACTA
  181 TAGGGAGAGC CGCCACCATG GCCCCAAAGA AGAAGCGGAA GGTCGGTATC CACGGAGTCC
  241 CAGCAGCCGA CAAGAAGTAC TCCATTGGGC TCGATATCGG CACAAACAGC GTCGGCTGGG
  301 CCGTCATTAC GGACGAGTAC AAGGTGCCGA GCAAAAAATT CAAAGTTCTG GGCAATACCG
  361 ATCGCCACAG CATAAAGAAG AACCTCATTG GCGCCCTCCT GTTCGACTCC GGGGAGACGG
  421 CCGAAGCCAC GCGGCTCAAA AGAACAGCAC GGCGCAGATA TACCCGCAGA AAGAATCGGA
  481 TCTGCTACCT GCAGGAGATC TTTAGTAATG AGATGGCTAA GGTGGATGAC TCTTTCTTCC
  541 ATAGGCTGGA GGAGTCCTTT TTGGTGGAGG AGGATAAAAA GCACGAGCGC CACCCAATCT
  601 TTGGCAATAT CGTGGACGAG GTGGCGTACC ATGAAAAGTA CCCAACCATA TATCATCTGA
  661 GGAAGAAGCT TGTAGACAGT ACTGATAAGG CTGACTTGCG GTTGATCTAT CTCGCGCTGG
  721 CGCATATGAT CAAATTTCGG GGACACTTCC TCATCGAGGG GGANCTGAAC CCAGACAACA
  781 GCGATGTCGA CAAACTCTTT ATCCNACTGG TTCAGACTTA CNATCAGCTT TTCGAANAGA
  841 ACCCGATCAA CGCATCCGGA NTTGANGCCN AAGCAATCCT GANCGCTANG CTGTCCAAAT
  901 CCCGGCGGNT CGAAAACCTC ATCGCACAGC TCCNNNGGGN NNNNAANNNN NNNCNGNTNG
961 GNAATNNTAT CNCCNNNNNN CTCGGGCNGA NCCCNNNCTT TNANNNNNNN NNN
//
Primer: HSV_TK_pA_R
Region: SV40 NLS, insert 3'
Sequence file: RDB13130_A5Bke.seq check
>13130_13130_A5Bk_HSV_tk_pA_R_3_A03_03.ab1
    1 NNNNNNNNNN NNNNNTTGNC NNNTTCCGNG TTTNNGTTAG CCNCCCCCGT TTAAACTCAT
   61 TACTAACCGG TAGGGATCGA ACCCTTTCAC ACCTTCCTCT TCTTCTTGGG GTCAGCCCTG
  121 CTGTCTCCAC CGAGCTGAGA GAGGTCGATT CTTGTTTCAT AGAGCCCCGT AATTGACTGA
  181 TGAATCAGTG TGGCGTCCAG GACCTCCTTT GTAGAGGTGT ACCGCTTTCT GTCTATGGTG
  241 GTGTCGAAGT ACTTGAAGGC TGCAGGCGCG CCCAAGTTGG TCAGAGTAAA CAAGTGGATA
  301 ATGTTTTCTG CCTGCTCCCT GATGGGCTTA TCCCTGTGCT TATTGTAAGC AGAAAGCACC
  361 TTATCGAGGT TAGCGTCGGC GAGGATCACT CTTTTGGAGA ATTCGCTTAT TTGCTCGATG
  421 ATCTCATCAA GGTAGTGTTT GTGTTGTTCC ACGAACAGCT GCTTCTGCTC ATTATCTTCG
  481 GGAGACCCTT TGAGCTTTTC ATAGTGGCTG GCCAGATACA AGAAATTAAC GTATTTAGAG
  541 GGCAGTGCCA GCTCGTTACC TTTCTGCAGC TCGCCCGCAC TAGCGAGCAT TCGTTTCCGG
  601 CCGTTTTCAA GCTCAAAGAG AGAGTACTTG GGAAGCTTAA TGATGAGGTC TTTTTTGACC
  661 TCTTTATATC CTTTCGCCTC GAGAAAGTCG ATGGGGTTTT TTTCGAAGCT TGATCGCTCC
  721 ATGATTGTGA TGCCCAGCAG TTCCTTGACG CTTTTGAGTT TTTTAGACTT CCCTTTCTCC
  781 NCTTTGGNCA CAACCNGTAC ACTGTAAGCG ACTGTAGGAG AATCGAATCC GCCGNATTTC
  841 TNGGGGTCCC NATCTTTTTT GCGNGCGATC AGCTNGNCGC NGTNNCTTTT CGGGANGANA
  901 CTTNNCTNNG ANAANNCTCC GGNCNGNACT TCGGNCTTTT TAACNATGNT NNCCNGCNGN
  961 NNNNNNNGNN NNNNNCNGNA NNNNCNNGNA NNCCNNACCN NTNNNNCNNN NNNNNTNNNN
1021 NNNNTNNNNN NNNNN
//
Primer: hCAS9+2010_F
Region: insert mid (H840 WT)
Sequence file: RDB13130_A5Bkf.seq check
>13130_13130_B1HiF_hCAS9+2010_F_G01_07_ABI08.ab1
    1 NNNNNNNNNN NNNNNTTNNN NNNNCCGANG GNNTTGCNNA CCGGAACTTC NTGCAGTTGA
   61 TCCATGATGA CTCTCTCACC TTTAAGGAGG ACATCCAGAA AGCACAAGTT TCTGGCCAGG
  121 GGGACAGTCT TCACGAGCAC ATCGCTAATC TTGCAGGTAG CCCAGCTATC AAAAAGGGAA
  181 TACTGCAGAC CGTTAAGGTC GTGGATGAAC TCGTCAAAGT AATGGGAAGG CATAAGCCCG
  241 AGAATATCGT TATCGAGATG GCCCGAGAGA ACCAAACTAC CCAGAAGGGA CAGAAGAACA
  301 GTAGGGAAAG GATGAAGAGG ATTGAAGAGG GTATAAAAGA ACTGGGGTCC CAAATCCTTA
  361 AGGAACACCC AGTTGAAAAC ACCCAGCTTC AGAATGAGAA GCTCTACCTG TACTACCTGC
  421 AGAACGGCAG GGACATGTAC GTGGATCAGG AACTGGACAT CAATCGGCTC TCCGACTACG
  481 ACGTGGATCA TATCGTGCCC CAGTCTTTTC TCAAAGATGA TTCTATTGAT AATAAAGTGT
  541 TGACAAGATC CGATAAAAAT AGAGGGAAGA GTGATAACGT CCCCTCAGAA GAAGTTGTCA
  601 AGANNATGAA AAATTATTGG CGGCAGCTGC TGAACGCCAA ACTGATCACA CAACGGAAGT
  661 TCGATAATCT GACTAAGGCT GAACGAGGTG GCCTGTCTGA GTTGGATAAA GCCGGCTTCA
  721 TCAAAAGGCA GCTTGTTGAG ACACGCCAGA TCACCAAGCA CGTGGCCCAA ATTCTCGATT
  781 CACGCATGAA CACCAAGTAC GATGAAAATG ACAAACTGAT TCGAGAGGTG AAAGTTATTA
  841 CTCTGAANTC TNAGCTGGTC TCAGANTNCA GAANGGACNT TCAGTTTTAT AANGTGAGAG
  901 AGATCAACAA TNNCNACCNT GCGCATGATG CCNACCNGAA NGCNNNGGTN NNNNNNGNNN
961 TNATNNAAAA ANNNNNCNNN NNNNNNN
//
Primer: hCAS9+3550_R
Region: insert mid (R1060,K1003 WT)
Sequence file: RDB13130_A5Bkg.seq check
>13130_13130_B1HiF_hCAS9+3550_R_H01_08_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNGNNNGN GNNGCCCNGC AGTTCCTTGN CGCTTTTGAG
   61 TTTTTTAGAC TTCCCTTTCT CCACTTTGGC CACAACCAGT ACACTGTAAG CGACTGTAGG
  121 AGAATCGAAT CCGCCGTATT TCTTGGGGTC CCAATCTTTT TTGCGTGCGA TCAGCTTGTC
  181 GCTGTTCCTT TTCGGGAGGA TACTTTCCTT GGAGAAGCCT CCGGTCTGTA CTTCGGTCTT
  241 TTTAACGATG TTCACCTGCG GCATGGACAG GACCTTCCGG ACTGTCGCGA AATCCCTACC
  301 CTTGTCCCAC ACGATTTCTC CTGTTTCTCC GTTTGTTTCG ATAAGTGGTC GCTTCCGAAT
  361 CTCTCCATTG GCCAGTGTAA TCTCGGTCTT GAAAAAATTC ATAATATTGC TGTAAAAGAA
  421 GTACTTAGCG GTGGCCTTGC CTATTTCCTG CTCAGACTTT GCGATCATTT TCCTAACATC
  481 GTACACTTTA TAGTCTCCGT AAACAAATTC AGATTCAAGC TTGGGATATT TTTTGATAAG
  541 TGCAGTGCCT ACCACTGCAT TCAGGTAGGC ATCATGCGCA TGGTGGTAAT TGTTGATCTC
  601 TCTCACCTTA TAAAACTGAA AGTCCTTTCT GAAATCTGAG ACCAGCTTAG ACTTCAGAGT
  661 AATAACTTTC ACCTCTCGAA TCAGTTTGTC ATTTTCATCG TACTTGGTGT TCATGCGTGA
  721 ATCGAGAATT TGGGCCACGT GCTTGGTGAT CTGGCGTGTC TCAACAAGCT GCCTTTTGAT
  781 GAANCCGGCT TTATCCAACT CANANAGGCC ACCTCGTTCA GCCTTANTCA GANTATCGAA
  841 CTNNCGTTGN GTGATCANTT NGNCGTTCAN CAGCTGCCGC CNATANTTTT TNCNTTTTCT
  901 TGNNNNCTTN NNCNGANGGG ANGTTNNCAC NNNNCCNNNN ANTTTTNATC NGNNNNNTNN
961 NNNNNNNN
//
Primer: SV40pro_F
Region: NeoR
Sequence file: RDB13130_A5Bkh.seq check
>13130_13130_A5Bk_SV40pro_F_3_B03_06.ab1
    1 NNNNNNNNNN NNNNNNNNGA NNNGTGANGN NGNTTTTTTG GAGGNCTNNG CTTTTGCNAA
   61 AAGCTCCCGG GAGCTTGTAT ATCCNTTTTC GGATCTGATC AAGAGACAGG ATGAGGATCG
  121 TTTCGCATGA TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG
  181 CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG
  241 CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT
  301 GAACTGCAGG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA
  361 GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG
  421 GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT
  481 GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA
  541 CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG
  601 GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGCGCATG
  661 CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG
  721 GAAAATGGNC GCTTTTCTGG ANTCATCGAC TGTGGCCGGC TGGGNGTGGC GGANCGCTAT
  781 CAGGACATAG CGTTGGNTAC CCGTGATANN GCTGAAGANC TTGGCGGCGA ANGGGCTGAN
  841 CGCTTCCTCG NGCTTTACNG NATCGCCGCT CCCNANTCGC AGCGCATCGC CNNCTATNGC
  901 CNTCTNGANN ANNNCTTCTG ANCGGGNNNN NNGGGTNNGC GAAATGANNN ANNNNNCGAN
961 NNCNNNNCNT GNNNTCNCNA NATTNCNANN NNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Yoshimi, K., ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes. Nat. Commun. 7: 10431 (2016). PMID 26786405.
reference Hashikawa, Y., Generation of Knockout Rabbits With X-linked Severe Combined Immunodeficiency (X-SCID) Using CRISPR/Cas9. Sci. Rep. 10 (1): 9957 (2020).
user report Mizuno-Iijima, S., Efficient production of large deletion and gene fragment knock-in mice mediated by genome editing with Cas9-mouse Cdt1 in mouse zygotes. Methods 191: 23-31 (2021). PMID 32334080.
user report Suzuki, K., Deletion of B-cell translocation gene 2 (BTG2) alters the responses of glial cells in white matter to chronic cerebral hypoperfusion. J. Neuroinflammation 18 (1): 86 (2021). PMID 33812385.
user report Osawa, Y., EXOC1 plays an integral role in spermatogonia pseudopod elongation and spermatocyte stable syncytium formation in mice. Elife 10: e59759 (2021). PMID 33973520.
user report Matsumura, K., Pathogenic POGZ mutation causes impaired cortical development and reversible autism-like phenotypes. Nat. Commun. 11 (1): 859 (2020). PMID 32103003.
user report Ayabe, S., Off- and on-target effects of genome editing in mouse embryos. J. Reprod. Dev. 65 (1): 1-5 (2019). PMID 30518723.
user report Miyasaka, Y., CLICK: one-step generation of conditional knockout mice. BMC Genomics 19 (1): 318 (2018). PMID 29720086.

2021.09.16

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