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

px330-mC (#RDB14406)

Expression vector of sgRNA with hSpCas9-Cdt1(mouse) fusion protein.

Clone info. Expression vector of sgRNA with hSpCas9-Cdt1(mouse) fusion protein (aa29-132, fused to Cas9 C-terminus).
Vector backbone pX330-U6-Chimeric_BB-CBh-hSpCas9 (plasmid)
Size of vector backbone 8.5 kb
Selectable markers Ampicillin
Growth conditions LB+Amp, 37oC
Gene/insert name mouse Cdt1 cDNA S. pyogenes Cas9 genomic
Depositor|Developer Sugiyama, Fumihiro | Mizuno, Seiya |
 

Distribution information

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Please visit Information of Request for Distribution.[link] 
Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Mizuno-Iijima, S. et al., Methods 191: 23-31 (2021)).
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.
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必要書類
提供依頼書  [依頼書の記入例 ]
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]
手続きの概要は、「提供申込みについて[link]」をご覧ください。
MTAに書く使用条件 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 (Mizuno-Iijima, S. et al., Methods 191: 23-31 (2021))。
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.)
RDB14406 px330-mC DNA solution

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

How to cite this biological resource

Materials & Methods section:

The px330-mC was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14406).

Reference section:

Mizuno-Iijima, S., Ayabe, S., Kato, K., Matoba, S., Ikeda, Y., Dinh, T.T.H., Le, H.T., Suzuki, H., Nakashima, K., Hasegawa, Y., Hamada, Y., Tanimoto, Y., Daitoku, Y., Iki, N., Ishida, M., Ibrahim, E.A.E., Nakashiba., , T., Hamada, M., Murata, K., Miwa, Y., Okada-Iwabu, M., Iwabu, M., Yagami, K., Ogura, A., Obata, Y., Takahashi, S., Mizuno, S., Yoshiki, A., Sugiyama, F., 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. [link to RRC of NBRP]

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

QC test results

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 RDB14406_A6L1p1-2.pdf check

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

Primer: SV40pA3element Reverse (Pr0108)
Region: U6 pro,gRNA scaffold
Sequence file: RDB14406_A6L1d.seq check
>D04199A2_A6L1_1_SV40pA3'element Reverse_D04_10_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNANNGN ANANNNCCNN NCNANCGCGG CCTTTTTACG
   61 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTGAGGG CCTATTTCCC ATGATTCCTT
  121 CATATTTGCA TATACGATAC AAGGCTGTTA GAGAGATAAT TGGAATTAAT TTGACTGTAA
  181 ACACAAAGAT ATTAGTACAA AATACGTGAC GTAGAAAGTA ATAATTTCTT GGGTAGTTTG
  241 CAGTTTTAAA ATTATGTTTT AAAATGGACT ATCATATGCT TACCGTAACT TGAAAGTATT
  301 TCGATTTCTT GGCTTTATAT ATCTTGTGGA AAGGACGAAA CACCGGGTCT TCGAGAAGAC
  361 CTGTTTTAGA GCTAGAAATA GCAAGTTAAA ATAAGGCTAG TCCGTTATCA ACTTGAAAAA
  421 GTGGCACCGA GTCGGTGCTT TTTTGTTTTA GAGCTAGAAA TAGCAAGTTA AAATAAGGCT
  481 AGTCCGTTTT TANNNCGTGN NNCNATTCTG CAGACAAATG GCTCTAGAGG TACCCGTTAC
  541 ATAACTTACG GTAAATGGCC CGCCTGGCTG ACCGCCCAAC GACCCCCGCC CATTGACGTC
  601 AATAGTAACG CCAATAGGGA CTTTCCATTG ACGTCAATGG GTGGAGTATT TACGGTAAAC
  661 TGCCCACTTG GCAGTACATC AAGTGTATCA TATGCCAAGT ACGCCCCCTA TTGACGTCAA
  721 TGACGGTAAA TGGCCCGCCT GGCATTGTGC CCAGTACATG ACCTTATGGN ANTTTCCTAC
  781 TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG GGNCGANGTG AGNCCCACGT
  841 TCTGCTNCAC TCTCCCCATC TCCCCCCCCC TCCCCCNNCC CCCAATTTTG NATTTATTTA
901 TTTTTTTANN TATTTTTGTG CANNNANNGG GGGCGGGGGG GGGGGGGGGG GN
//
Primer: RDB13535#1 (Pr0302)
Region: 3XFLAG,SV40 NLS,Cas9 5'
Sequence file: RDB14406_A6L1e.seq check
>D04199A2_A6L1_1_RDB13535#1_E04_13_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNTTT TTTTNNGGTT GGNNCGGTGC CNCCATGGNC TATAAGGACC
   61 ACGACGGAGA CTACAAGGAT CATGATATTG ATTACAAAGA CGATGACGAT AAGATGGCCC
  121 CAAAGAAGAA GCGGAAGGTC GGTATCCACG GAGTCCCAGC AGCCGACAAG AAGTACAGCA
  181 TCGGCCTGGA CATCGGCACC AACTCTGTGG GCTGGGCCGT GATCACCGAC GAGTACAAGG
  241 TGCCCAGCAA GAAATTCAAG GTGCTGGGCA ACACCGACCG GCACAGCATC AAGAAGAACC
  301 TGATCGGAGC CCTGCTGTTC GACAGCGGCG AAACAGCCGA GGCCACCCGG CTGAAGAGAA
  361 CCGCCAGAAG AAGATACACC AGACGGAAGA ACCGGATCTG CTATCTGCAA GAGATCTTCA
  421 GCAACGAGAT GGCCAAGGTG GACGACAGCT TCTTCCACAG ACTGGAAGAG TCCTTCCTGG
  481 TGGAAGAGGA TAAGAAGCAC GAGCGGCACC CCATCTTCGG CAACATCGTG GACGAGGTGG
  541 CCTACCACGA GAAGTACCCC ACCATCTACC ACCTGAGAAA GAAACTGGTG GACAGCACCG
  601 ACNAGGCCGA CCTGCGGCTG ATCTATCTGG CCCTGGCCCA CATGATCAAG TTCCGGGGCC
  661 ACTTCCTGAT CGAGGGCGAN CTGAACCCCG ACAACAGCGA CGTGGACAAG CTGTTCATCC
  721 AGCTGGTGCA GACCTACAAC CAGCTGTTCG ANGAAAACCN CATCAACGCC NGCGGCGTGG
  781 ANGCCNAGGC CATCCTGTCT GCCNGANTGA NCAAGANCAG ANGGCTGGAA AATCTGATCG
  841 CCCAGCTGCC CGGCGANAAG AANAATGGNC NGTNCGGAAA NCNGATTGNC NNGAGCCNGG
  901 GNNNGACCCC CNNCTTNNNN ANCNNCTTCN ANNNNNNNNN ANNNNGNNNA NNTGNAGCNN
961 ANCNANNNAN NNCNTACNNN NANNANCNNN NNNNCNNNNN NGNNNNNNNN NN
//
Primer: BGH_rev2 (Pr0606)
Region: bGHpA,SV40 NLS,mCdt1,Cas9 3'
Sequence file: RDB14406_A6L1f.seq check
>D04199A2_A6L1_1_BGH_rev2_F04_16_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNGGG GGNNGGNNNN NNACAGATGG CTGGCAACTA GAAGGCACAG
   61 TCGAGGCTGA TCAGCGAGCT CTAGGAATTC TTAGCTGGCC TCCACCTTTC TCTTCTTCTT
  121 GGGGCTGAAT TCGACGGAGT CCTCTGAGGG GATCTTGCCC GGCTGTTGAC CAACATAAAC
  181 AGTTGTACTC TTTGTTCGCT TGACGGGGGA TGGGCAAACG GGGGAGCCAG GGTCAGCTGG
  241 AGGGCTAGGG TCAGCTGGAG GGCTGGGCTC AGCTGGGGAA CTGGGCTCAG GCAGAGAACT
  301 GGGGCAGGAG TCCAATCCAG GCAGCCGTAA CCTCCGGCGC GCGAGCGGCG CGGGCTGGTC
  361 ACTCCCGGGT TCGGCTGGGG GCCGGGCGCG CTTGCGGCTG CTGCTCCGGG TGAACGCAGG
  421 AGCCACGAGG CCACCAGGGC TCGGGTCGCC TCCCAGCTGA GACAGGTCGA TCCGTGTCTC
  481 GTACAGGCCG GTGATGCTCT GGTGGATCAG GGTGGCGTCC AGCACCTCTT TGGTGCTGGT
  541 GTACCTCTTC CGGTCGATGG TGGTGTCAAA GTACTTGAAG GCGGCAGGGG CTCCCAGATT
  601 GGTCAGGGTA AACAGGTGGA TGATATTCTC GGCCTGCTCT CTGATGGGCT TATCCCGGTG
  661 CTTGTTGTAG GCGGACAGCA CTTTGTCCAG ATTAGCGTCG GCCAGGATCA CTCTCTTGGA
  721 NAACTCGCTG ATCTGCTCGA TGANCTCGTC CAGGTAGTGC TTGTGCTGTT CCACAAACAG
  781 CTGTTTCTGC TCATTATCCT CGGGGGANCC NNTCAGCTTC TCATAGTGGC TGGNCAGGTN
  841 CAGGAANNTC ACATATTTGG ANGGNANGGN CAGTTCGTTT CCNNTCTGCA GTTCGCCNGC
  901 ANANGNNANC ATTNNCTTCC NGNNNNTTTC CNNNNCNNAN NNNGNNNNAC NTNNNNAGCT
961 NNNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original 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. [link to RRC of NBRP]
reference Fujita, R., Generation of a MyoD knock-in reporter mouse line to study muscle stem cell dynamics and heterogeneity. iScience 26 (5): 106592 (2023). PMID 37250337.