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pAAV2-TRE-mito-SRAI-BGHpA

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

Catalog number RDB18684
Resource name pAAV2-TRE-mito-SRAI-BGHpA
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
Gene/insert name Aequorea victoria YPet (GFP) cDNA Galaxea fascicularis TOLLES (GFP) cDNA
Depositor|Developer Miyawaki, Atsushi | Hioki, Hiroyuki | Katayama, Hiroyuki |
Other clones in our bank

External Database
Aequorea victoria YPet (GFP)
    Galaxea fascicularis TOLLES (GFP)

            Reference sequence
              
             
            Sequence (full) RDB18684zzk02.seq provided by the depositor

            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 for distribution 1. The RECIPIENT of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR.
            2. A RECIPIENT who belongs to a non-profit organization may use the BIOLOGICAL RESOURCE for an academic research that is non-commercial.
            3. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR or an acknowledgment to the DEPOSITOR are 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. (2020). Cell 181, 1176–1187.
            4. The RECIPIENT agrees to provide a copy of the publication to the DEPOSITOR, oral or written, reporting use of the BIOLOGICAL RESOURCE.
            5. The Recipient’s Scientist agrees not to transfer the BIOLOGICAL RESOURCE to any third party including any resource centers without the prior written consent of DEPOSITOR.
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            MTA, for use for not-for-profit academic purpose [Word]
            Approval form [Word] by Dr. Atsushi Miyawaki
            Please visit Information of Request for Distribution.[link] 
            Remarks Remember that you will be working with samples containing infectious virus.
            Please obtain written approval of Dr. Atsushi Miyawaki.
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            提供条件 1. The RECIPIENT of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR.
            2. A RECIPIENT who belongs to a non-profit organization may use the BIOLOGICAL RESOURCE for an academic research that is non-commercial.
            3. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR or an acknowledgment to the DEPOSITOR are 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. (2020). Cell 181, 1176–1187.
            4. The RECIPIENT agrees to provide a copy of the publication to the DEPOSITOR, oral or written, reporting use of the BIOLOGICAL RESOURCE.
            5. The Recipient’s Scientist agrees not to transfer the BIOLOGICAL RESOURCE to any third party including any resource centers without the prior written consent of DEPOSITOR.
            提供依頼
            提供依頼書 [Word]
            提供同意書 (MTA, 非営利学術目的用)[Word]
            宮脇敦史博士による提供承諾書 [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。
            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。
            宮脇敦史博士の書面による提供承諾が必要です。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB18684 pAAV2-TRE-mito-SRAI-BGHpA 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 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)


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            Featured content

            Featured content Mitophagy visualization fluorescent sensor, mito-SRAI (English text)
            Featured content マイトファジー可視化蛍光センサー mito-SRAI (Japanese text)
            Featured content Mitophagy (English text)

            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]

            2024.03.26

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