RDB18684 - pAAV2-TRE-mito-SRAI-BGHpA

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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

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.

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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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Ordering forms	Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ] MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ] Please visit Information of Request for Distribution.[link]
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.

必要書類	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

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

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
>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
>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
>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
>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
>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
>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]

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Other resources in our bank

External Database
Aequorea victoria YPet (GFP)

Galaxea fascicularis TOLLES (GFP)

Reference sequence