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pAAV2-TRE-mito-SRAI-BGHpA (#RDB18684)

AAV2 expression vector of mito-SRAI (signal-retaining autophagy indicator) driven by the tetracyclin-responsive element promoter for quantitative visualizing mitophagy.

Clone info. AAV2 expression vector of mito-SRAI (signal-retaining autophagy indicator) driven by the tetracyclin-responsive element (TRE-tight) promoter. mito-SRAI is a fluorescent sensor for quantitative visualizing mitophagy. SRAI is a tandem fusion of YPet and TOLLES, fused with a DNA linker that encodes a triple repeat of the amino acid linker Gly-Gly-Gly-Gly-Ser. The ITRs (inverted terminal repeats) are derived from AAV serotype 2.
Comment Expression was confirmed by the depositor, co-transfection to cultured cell with a plasmid vector that expresses tTA under ubiquitous promoter.
Vector backbone plasmid
Selectable markers Ampicillin
Growth conditions LB+Amp, 37oC
Gene/insert name Aequorea victoria YPet (GFP) cDNA Galaxea fascicularis TOLLES (GFP) cDNA
Depositor|Developer Miyawaki, Atsushi | Hioki, Hiroyuki | Katayama, Hiroyuki |
 
Sequence (full) RDB18684zzk02.seq provided by the depositor

Distribution information

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Terms and conditions for distribution 1. If the BIOLOGICAL RESOURCE is to be used by a for-profit organization, the RECIPIENT must agree with RIKEN Innovation Co., Ltd. on the terms and conditions of its use. For inquiries, please contact:
license-contact@innovation-riken.jp
2. The RECIPIENT’s Scientist agrees not to transfer the BIOLOGICAL RESOURCE and its derivatives to any third party including any resource center without the prior written consent of DEPOSITOR.
3. When publishing the research results obtained using the BIOLOGICAL RESOURCE, a citation to the literature specified by the DEPOSITOR or an acknowledgement to the DEPOSITOR is requested.
Sohn, J. et al., PLoS One, 12 (1): e0169611 (2017). Hioki, H. et al., Neurosci. Res., 63 (2): 149-154 (2009). Katayama et al. Cell 181, 1176–1187 (2020).
Remarks Remember that you will be working with samples containing infectious virus.
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提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
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 1. If the BIOLOGICAL RESOURCE is to be used by a for-profit organization, the RECIPIENT must agree with RIKEN Innovation Co., Ltd. on the terms and conditions of its use. For inquiries, please contact:
license-contact@innovation-riken.jp
2. The RECIPIENT’s Scientist agrees not to transfer the BIOLOGICAL RESOURCE and its derivatives to any third party including any resource center without the prior written consent of DEPOSITOR.
3. When publishing the research results obtained using the BIOLOGICAL RESOURCE, a citation to the literature specified by the DEPOSITOR or an acknowledgement to the DEPOSITOR is requested.
Sohn, J. et al., PLoS One, 12 (1): e0169611 (2017). Hioki, H. et al., Neurosci. Res., 63 (2): 149-154 (2009). Katayama et al. Cell 181, 1176–1187 (2020).
備考 このリソースはウイルス粒子産生用のため、取扱いに注意が必要です。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB18684 pAAV2-TRE-mito-SRAI-BGHpA DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge

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 pAAV2-TRE-mito-SRAI-BGHpA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB18684).

Reference section:

Katayama, H., Hama, H., Nagasawa, K., Kurokawa, H., Sugiyama, M., Ando, R., Funata, M., Yoshida, N., Homma, M., Nishimura, T., Takahashi, M., Ishida, Y., Hioki, H., Tsujihata, Y., Miyawaki, A., Visualizing and Modulating Mitophagy for Therapeutic Studies of Neurodegeneration. Cell 181(5): 1176-1187 (2020). PMID 32437660. [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 RDB18684_B1B5p1-1.pdf check

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

Primer: EGFP-N (Pr0027)
Region: mito-SRAI 5' (2xCOX8 pre seq), tight TRE pro, attB2, AAV2 ITR
Sequence file: RDB18684_B1B5a.seq check
>D07612A1_B1B5_1_EGFP-N_A01_01_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNTCGCCCTT GCTNNNCNTG GNNCCCCCGA NCTTCCCCTC
   61 CGGCGGCAAC GAATGGATCT TGGCGCGCGG CACTGGGAGC CGCCGGGCCG AGCCTGTCAA
  121 GCCCCGCAGC AGCAGCGGCG TCAGGACGGA CATCCCAAGC TTCCCCTCCG GCGGCAACGA
  181 ATGGATCTTG GCGCGCGGCA CTGGGAGCCG CCGGGCCGAG CCTGTCAAGC CCCGCAGCAG
  241 CAGCGGCGTC AGGACGGACA TGGTGGTACC GAGCTCGAAT TCTCCAGGCG ATCTGACGGT
  301 TCACTAAACG AGCTCTGCTT ATATAGGCCT CCCACCGTAC ACGCCTACCT CGACATACGT
  361 TCTCTATCAC TGATAGGGAG TAAACTCGAC ATACGTTCTC TATCACTGAT AGGGATAAAC
  421 TCGACATACG TTCTCTATCA CTGATAGGGA GTAAACTCGA CATACGTTCT CTATCACTGA
  481 TAGGGAGTAA ACTCGACATA CGTTCTCTAT CACTGATAGG GAGTAAACTC GACATCGTTC
  541 TCTATCACTG ATAGGGAGTA AACTCGACAT ACGTTCTCTA TCACTGATAG GGAGTAAACT
  601 CGAGATATCT AGACCCAGCT TTCTTGTACA AAGTGGTACG CGTGCGGCCG CAGGAACCCC
661 TAGTGATGGA GTTGGCCACT CCCTCTCTGC GCGCTN
//
Primer: BGHpA_F (Pr0373)
Region: bGH polyA, attB1, AAV2 ITR
Sequence file: RDB18684_B1B5b.seq check
>D07612A1_B1B5_1_BGHpA_F_E05_05_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNG GGGTGGGGCA GGNNAGCAAG GGGGANGNNT
   61 GGGAAGANAA TAGCAGGCAT GCTGGGGATG CGGTGGGCTC TATGGGCGGC CGCATCGATA
  121 AGCTTGTCGA CGATAAAGCC TGCTTTTTTG TACAAACTTG TCACGTGCGG ACCGAGCGGC
181 CGCAGGAACC CCTAGTGATG GAGTTGGCCA CTCCCTCTCT GCGCGCTN
//
Primer: CSII_CMV5_v2 (Pr0255)
Region: mito-SRAI mid 5' (2xCOX8 pre seq, EYFP)
Sequence file: RDB18684_B1B5c.seq check
>D07612A1_B1B5_1_CSII-CMV5_V2_C01_03_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNTNNCCG TCCTGACGCC GCTGCTGCTG
   61 CGGGGCTTGA CAGGCTCGGC CCGGCGGCTC CCAGTGCCGC GCGCCAAGAT CCATTCGTTG
  121 CCGCCGGAGG GGAAGCTTGG GATGTCCGTC CTGACGCCGC TGCTGCTGCG GGGCTTGACA
  181 GGCTCGGCCC GGCGGCTCCC AGTGCCGCGC GCCAAGATCC ATTCGTTGCC GCCGGAGGGG
  241 AAGCTCGGGG GATCCATGGT GAGCAAGGGC GAGGAGCTGT TCACCGGGGT GGTGCCCATC
  301 CTGGTCGAGC TGGACGGCGA CGTAAACGGC CACAAGTTCA GCGTGTCCGG CGAGGGCGAG
  361 GGCGATGCCA CCTACGGCAA GCTGACCCTG AAGCTGCTCT GCACCACCGG CAAGCTGCCC
  421 GTGCCCTGGC CCACCCTCGT GACCACCCTG GGCTACGGCC TGCAGTGCTT CGCCCGCTAC
  481 CCCGACCACA TGAAGCAGCA CGACTTCTTC AAGTCCGCCA TGCCCGAAGG CTACGTCCAG
  541 GAGCGCACCA TCTTCTTCAA GGACGACGGC AACTACAAGA CCCGCGCCGA GGTGAAGTTC
  601 GAGGGCGACA CCCTGGTGAA CCGCATCGAG CTGAAGGGCA TCGACTTCAA GGAGGACGGC
  661 AACATCCTGG GGCACAAGCT GGAGTACAAC TACAACAGCC ACAACGTCTA TATCACCGCC
  721 GANNAAGCAG AAGAACGGCA TCAAGGCCNA CTTCNAGATC CGCCACNACA TCGAGGANGG
  781 CGNNNNGNAG CTCGCCGACC ACTACCAGCA NANCACCCNC NTCGNNGANG GCCCCGTGCT
  841 GCTGCCCGAC AACCACTACC TGANCTACNA GTCCGNCCTG TTCNAAGANC CCCNACNANN
  901 AAGCGCGATC ACNNNGNNCC TGCTGGANTN NCTGACCGCN NNCNGGGATC ACNNGAANGC
  961 ATGAACNANC TGTNNAANGG ANCTNNGNNG GNANGCNGNT NNNNNNNGAN NTGGNNTCNN
 1021 NGNNGNNGGN NNNNNNGNNT CNANNANNGN NNNANCNNNN NNNAANNNNN NNNANNNNAN
1081 NNNTNANNCN NNNNNNNNN
//
Primer: mTurquoise2-M_F (Pr0577)
Region: mito-SRAI mid (EYFP, hmAzami-Green)
Sequence file: RDB18684_B1B5d.seq check
>D07612A1_B1B5_1_mTurquoise2-M_F_D01_04_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNTACCNG NNGAACACCC CCATCGGCGA
   61 CGGCCCCGTG CTGCTGCCCG ACAACCACTA CCTGAGCTAC CAGTCCGCCC TGTTCAAAGA
  121 CCCCAACGAG AAGCGCGATC ACATGGTCCT GCTGGAGTTC CTGACCGCCG CCGGGATCAC
  181 TGAAGGCATG AACGAGCTGT ACAAGGAGCT CGGTGGAGGC GGTTCAGGCG GAGGTGGCTC
  241 TGGCGGTGGC GGATCGCTCG AGATGGTGAG CGTGATCAAG CCCGAGATGA AGATCAAGCT
  301 GTGCATGAGG GGCACCGTGA ACGGCCACAA CTTCGTGATT GAGGGCGAGG GCAAGGGCAA
  361 CCCCTACGAG GGCACCCAGA TCCTGGACCT GAACGTGACC GAGGGCGCCC CCCTGCCCTT
  421 CGCCTACGAC ATCCTGTCCA CCTCGTTCCA GTACGGCAAC AGGGCCTTCA CCAAGTACCC
  481 CGCCGACATC CAGGACTACT TCAAGCAGGC CTTCCCCGAG GGCTACCACT GGGAGAGGAG
  541 CATGACCTAC GAGGACCAGG GCATCTGCAC CGCCACCAGC AACATCAGCA TGAGGGGCGA
  601 CTGCTTCTTC TACGACATCA GGTTCGACGG CACCAACTTC CCCCCCAACG GCCCCGTGAT
  661 GCAGAAGAAG ACCCTGAAGT GGGAGCCCGA CACCGAGAAG ATGTACGTGG AGGANGGCGT
  721 GCTGAAGGGC GACGTGAACA TGAGGCTGCT GCTGGAGGGC GGCGGCCACT ACAGGTGCGA
  781 CGTCNAGACC ACCTACAAGG CCAAGAANGA NGTGAGCCTG CCCGACGCCC ACAAGATCGA
  841 CCACAGGATC GAGATCCTGG ANCACGANNG GACTACAACA AGGTGANNNT GTGCGAGAAC
  901 GNCGNGNNCN GGGCCTCNNT GCTGCCCANC NNNGNCNANG CTANAGCCNG NNNANCNNGN
  961 TTCANCAGCC TGANNNATTT NNTGATNCNN NNNGNNAGNT CACNGNNNNN NGNGNNNACN
1021 NNNGNAANCN NANNNNNNNN NN
//
Primer: EGFP-C-rev (Pr0026)
Region: mito-SRAI mid (EYFP, COX8 pre seq)
Sequence file: RDB18684_B1B5e.seq check
>D07612A1_B1B5_1_EGFP-C-rev_B04_02_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTTNNNNNNN NNNGNNTGGT AGCTCAGGNN NGTGGTTGTC
   61 GGGCAGCAGC ACGGGGCCGT CGCCGATGGG GGTGTTCTGC TGGTAGTGGT CGGCGAGCTG
  121 CACGCCGCCG TCCTCGATGT TGTGGCGGAT CTTGAAGTTG GCCTTGATGC CGTTCTTCTG
  181 CTTGTCGGCG GTGATATAGA CGTTGTGGCT GTTGTAGTTG TACTCCAGCT TGTGCCCCAG
  241 GATGTTGCCG TCCTCCTTGA AGTCGATGCC CTTCAGCTCG ATGCGGTTCA CCAGGGTGTC
  301 GCCCTCGAAC TTCACCTCGG CGCGGGTCTT GTAGTTGCCG TCGTCCTTGA AGAAGATGGT
  361 GCGCTCCTGG ACGTAGCCTT CGGGCATGGC GGACTTGAAG AAGTCGTGCT GCTTCATGTG
  421 GTCGGGGTAG CGGGCGAAGC ACTGCAGGCC GTAGCCCAGG GTGGTCACGA GGGTGGGCCA
  481 GGGCACGGGC AGCTTGCCGG TGGTGCAGAG CAGCTTCAGG GTCAGCTTGC CGTAGGTGGC
  541 ATCGCCCTCG CCCTCGCCGG ACACGCTGAA CTTGTGGCCG TTTACGTCGC CGTCCAGCTC
  601 GACCAGGATG GGCACCACCC CGGTGAACAG CTCCTCGCCC TTGCTCACCA TGGATCCCCC
  661 GAGCTTCCCC TCCGGCGGCA ACGAATGGAT CTTGGCGCGC GGCACTGGGA GCCGCCGGGC
  721 CGAGCCTGTC AAGCCCCGCA GCAGCAGCGG CGTCAGGACG GACATCCCAN GCTTCCCCTC
  781 CGGCGGCAAC GAATGGATCT TGGCGCGCGG CACTGGGAGC CGCCGGGCCG ANCCTGTCAN
  841 GCCCCGCANC ANCAGCGGCG TCAGGACGGA CATGGNGNTA CCGANCTCGN ANTTCTCCNN
  901 GCGATCTGAC GGNTCACTAA ACGAGCTCTG CTTATATAGG CCTNCCNNCG TACACGCCTA
  961 CNNNNNCATA CGNTNNTCTA TNACTGATNN GGNNNNNNAA CNNGNNNNNN CNNTNNTCNA
1021 NNNNNGNNNN NGNNNNNNAA ANNTNNNNNN NNNN
//
Primer: bGH_rev3 (Pr0885)
Region: mito-SRAI 3' (hmAzami-Green)
Sequence file: RDB18684_B1B5f.seq check
>D07612A1_B1B5_1_bGH_rev3_B01_02_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGNN NGGCNNNNNN NNAGNTGGCT GGCNAACTAG
   61 AAGGCACNNN NNNGTCAGCT GACTNNAGGA NCCTACACAT TGATCCTAGC AGAAGCACAG
  121 GCTGCAGGGT GACGGTCCAT CCCGCTCTCC TGGGCACAAG ACATGGGCAG CGTGCCATCA
  181 TCCTGCTCCT CCACCTCCGG CGGGAAGCCA TGGCTCTGGA TCTGTTTCAT GAGTTGCCAC
  241 ATTGGCCGTG ATCTAGCCAG GTGGATCACG AAATGGCTCA GGCTGCTGAA CCAGTTCTTG
  301 CAGGCTCTAG CCTTGGCCTG GCTGGGCAGC ATGGAGGCCC TGGCCACGGC GTTCTCGCAC
  361 AGCTTCACCT TGTTGTAGTC CTTGTCGTGC TCCAGGATCT CGATCCTGTG GTCGATCTTG
  421 TGGGCGTCGG GCAGGCTCAC CTCCTTCTTG GCCTTGTAGG TGGTCTTGAC GTCGCACCTG
  481 TAGTGGCCGC CGCCCTCCAG CAGCAGCCTC ATGTTCACGT CGCCCTTCAG CACGCCGTCC
  541 TCCACGTACA TCTTCTCGGT GTCGGGCTCC CACTTCAGGG TCTTCTTCTG CATCACGGGG
  601 CCGTTGGGGG GGAAGTTGGT GCCGTCGAAC CTGATGTCGT AGAAGAAGCA GTCGCCCCTC
  661 ATGCTGATGT TGCTGGTGGC GGTGCAGATG CCCTGGTCCT CGTANGNNNN GNTCCTCTCC
  721 CNGTGGTAGC CCTCGGGGAA NGCCTGCTTG AAGTAGTCCT GNATGTCGGC GGGGTACTTG
  781 GTGAAGGCCC TGTTGCCGTA CTGNANCGAG GTGNNNAGGA TGTCGTANGC GAANNGNNAG
  841 GGGGGCGCCC TCNGTNNCGT NCANGNNNNN GATCTGGGNG CCCTCGTANG GNNNGCCCNT
  901 GCCCTCNCCC TCAANCNCNN NNNNGTGGCC GTTCACNGNN GCCCCTNNNG NNNNNCTTGA
  961 TNNTNNNTCN CNGGNNNNNT CNNGCTCACN NTCNNNNNNN CNANNCNNNC NNNNNCCNNN
1021 ANCNNNNCNN CCNNNNNGNA ANNNNCNNNN NNCNNNNNNN NNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Katayama, H., Visualizing and Modulating Mitophagy for Therapeutic Studies of Neurodegeneration. Cell 181(5): 1176-1187 (2020). PMID 32437660. [link to RRC of NBRP]