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Please review the QC test results indicated by check icon below as well as clone information before placing your order.

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.
Comment 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

External Database
Streptococcus pyogenes Cas9

          Reference sequence
            

          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.
          Ordering Please visit Information of Request for Distribution.[link] 
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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 contains CRISPR/Cas9 technologies and is not provided to for-profit organization or for for-profit research by non-profit organizations.
          提供案内 (日本国内) [open/close]

          提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Yoshimi, K. et al. Nat. Commun. 7: 10431, 2016)。非営利機関が非営利目的の教育・研究用に用いる場合以外は、大阪大学と別途MTAを締結すること。
          提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。
          提供依頼書 [Word]
          専用MTA(CRISPR/Cas9内包遺伝子材料専用 非営利学術目的)をお使いください [Word]
          備考
          本件リソースはCRISPR/Cas9 technologyを用いたゲノム編集バイオリソースです。営利機関および非営利機関による営利目的研究には提供いたしません。

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB13130 T7-NLS hCas9-pA DNA solution

          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.

          2022.05.17

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