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AAV:NT-probe-nuc-mCherry (#RDB19166)

Adeno-associated viral vector encoding the dGRAPHIC NT probe.

Clone info. Adeno-associated viral (AAV-Dj/8) vector encoding the dGRAPHIC (diffusible GPI anchored Reconstitution-Activated Proteins Highlight Intercellular Connections) NT probe (split sites with the 1–7 N terminal) fragment. The nucleic-localized mCherry is fused to the NT probe through a self-cleavable T2A peptide. This clone is used in conjunction with AAV:CT-probe-cyto-mCherry (RDB19167) as a tool to visualize the shape of cells after cell-cell contact. For more information, see Kinoshita, N. et al. Sci. Rep., 10 (1): 14437, 2020.
Comment Expression was confirmed by the depositor with confocal imaging.
Vector backbone pAAV-MCS (plasmid)
Size of vector backbone 7.3 kb
Selectable markers Ampicillin (E. coli)
Gene/insert name mouse Acr cDNA mouse Thy1 cDNA human histone H2B cDNA
Depositor|Developer Shimogori, Tomomi | Kinoshita, Nagatoki |
 
Sequence (partial) RDB19166pka01.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.
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 In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Kinoshita, N. et al., Sci. Rep., 10 (1): 14437, 2020).
Remarks Remember that you will be working with samples containing infectious virus.
This RESOURCE contains a fluorescent protein provided by Prof. Roger Tsien. UCSD's MTA is also required. Please visit Additional Forms section. This RESOURCE 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 ]
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 利用者は、研究成果の公表にあたって譲渡者の指定する文献を引用する (Kinoshita, N. et al., Sci. Rep., 10 (1): 14437, 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。
Prof. Roger Tsienから供与された蛍光タンパク質を含みます。UCSDとのMTA締結が必要です。「申込みに必要な書式」をご覧ください。本件リソースは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.)
RDB19166 AAV:NT-probe-nuc-mCherry 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 AAV:NT-probe-nuc-mCherry was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB19166).

Reference section:

Kinoshita, N., Huang, A.J.Y., McHugh, T.J., Miyawaki, A., Shimogori, T., Diffusible GRAPHIC to visualize morphology of cells after specific cell-cell contact. Sci. Rep. 10 (1): 14437 (2020). PMID 32879377. [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 RDB19166_B1F2p1-1.pdf check

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

Primer: EF1a-C_F (Pr0168)
Region: EF-1a pro, insert 5'
Sequence file: RDB19166_B1F2a.seq check
>D07910A1_B1F2_1_EF1a-C_F_D10_10_ABI24.ab1
    1 NNNNNNNNNN NNNTTTTGAN TTNGGNNCTT GGTTCNTTCT CAAGCCTCAG ACAGTGGTTC
   61 NAAGTTTTTT TCTTCCNTTT CAGGTGTCGT GAGGTACCGG ATCCTCTAGA CACCATGGTA
  121 GAGATGCTGC CAACTGTCGC TGTGCTGGTC TTGGCAGTGT CCGTGGTTGC CAAGGATAAC
  181 ACCACGCTGC AGGCTAGAGT GAGCAAGGGC GAGGAGCTGT TCACCGGGGT GGTGCCCATC
  241 CTGGTCGAGC TGGACGGCGA CGTAAACGGC CACAAGTTCA GCGTGCGCGG CGAGGGCGAG
  301 GGCGATGCCA CCATCGGCAA GCTGACCCTG AAGTTCATCT GCACCACCGG CAAGCTGCCC
  361 GTGCCCTGGC CCACCCTCGT GACCACCCTG ACCTACGGCG TGCAGTGCTT CAGCCGCTAC
  421 CCCGACCACA TGAAGCAGCA CGACTTCTTC AAGTCCGCCA TGCCCGAAGG CTACGTCCAG
  481 GAGCGCACCA TCTCCTTCAA GGACGACGGC AAATACAAGA CCCGCGCCGT TGTGAAGTTC
  541 GAGGGCGACA CCCTGGTGAA CCGCATCGAG CTGAAGGGCA CCGACTTCAA GGAGGACGGC
  601 AACATCCTGG GGCACAAGCT GGAGTACAAC TTCAACAGCC ACAACGTCTA TATCACGGCC
  661 GACAAGCAGG CTAGCTACCA ACTCGAGAAT GGCGGCATAA GCCTGCTGGT TCAGAACACA
  721 TCCTGGATGC TGCTGCTGCT GCTTTCCCTC TCCCTCCTCC AAGCCCTGGA CTTCATTTCT
  781 CTGGCATGCG AGGGCAGANG AANTCTTCTA ACATGCGGTG ACGTCNANGA NAATCCTGNN
  841 CNANTGCCAG ANCCAGCGAA NTCTGCTCCC GCCCCGAAAA ANGGCTCCNA GAANGCGGNN
901 NNNNNNNN
//
Primer: mCherry-check_R5 (Pr1024)
Region: insert 3'
Sequence file: RDB19166_B1F2b.seq check
>D07910A1_B1F2_1_mCherry-check_R5_C01_03_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNGNNNCCNC NTAGCGTNNA GGAGCAACAT AGTTAAGAAT
   61 ACCAGTCAAT CTTTCACAAA TTTTGTAATC CAGAGGTTGA TTATCGATAA GCTTGATATC
  121 CTATTACTTG TACAGCTCGT CCATGCCGCC GGTGGAGTGG CGGCCCTCGG CGCGTTCGTA
  181 CTGTTCCACG ATGGTGTAGT CCTCGTTGTG GGAGGTGATG TCCAACTTGA TGTTGACGTT
  241 GTAGGCGCCG GGCAGCTGCA CGGGCTTCTT GGCCTTGTAG GTGGTCTTGA CCTCAGCGTC
  301 GTAGTGGCCG CCGTCCTTCA GCTTCAGCCT CTGCTTGATC TCGCCCTTCA GGGCGCCGTC
  361 CTCGGGGTAC ATCCGCTCGG AGGAGGCCTC CCAGCCCATG GTCTTCTTCT GCATTACGGG
  421 GCCGTCGGAG GGGAAGTTGG TGCCGCGCAG CTTCACCTTG TAGATGAACT CGCCGTCCTG
  481 CAGGGAGGAG TCCTGGGTCA CGGTCACCAC GCCGCCGTCC TCGAAGTTCA TCACGCGCTC
  541 CCACTTGAAG CCCTCGGGGA AGGACAGCTT CAAGTAGTCG GGGATGTCGG CGGGGTGCTT
  601 CACGTAGGCC TTGGAGCCGT ACATGAACTG AGGGGACAGG ATGTCCCAGG CGAAGGGCAG
  661 GGGGCCACCC TTGGTCACCT TCAGCTTGGC GGTCTGGGTG CCCTCGTAGG GGCGGCCCTC
  721 GCCCTCGCCC TCGATCTCGA ACTCGTGGCC GTTCACGGAG CCCTCCATGT GCACCTTGAA
  781 NCGCATGAAC TCCTTGATGA TGGCCATGTT ATCCTCCTCG CCCTTGCTCA CCATGGTGGC
  841 TGGATNCTTA NCGCTGGTGN ACTTGGTGAT GGCCTTAGNA NCCTCNGANA CNGCGNGCTN
  901 GGCCNACTCC CCNNNNANCA NCANGNGCAC NGNNNNNNNG GNTCTNCCNN NNNGTGANGG
961 NCGANCGCTN NNNGNNNNNN NNNN
//
Primer: VN_F1 (Pr0992)
Region: insert mid
Sequence file: RDB19166_B1F2c.seq check
>D07910A1_B1F2_1_VN_F1_F10_16_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNC GTCTATNTNN NNGCCGNNNA GCAGGCTAGC
   61 TACCAACTCG AGAATGGCGG CATAAGCCTG CTGGTTCAGA ACACATCCTG GATGCTGCTG
  121 CTGCTGCTTT CCCTCTCCCT CCTCCAAGCC CTGGACTTCA TTTCTCTGGC ATGCGAGGGC
  181 AGAGGAAGTC TTCTAACATG CGGTGACGTC GAGGAGAATC CTGGCCCAAT GCCAGAGCCA
  241 GCGAAGTCTG CTCCCGCCCC GAAAAAGGGC TCCAAGAAGG CGGTGACTAA GGCGCAGAAG
  301 AAAGGCGGCA AGAAGCGCAA GCGCAGCCGC AAGGAGAGCT ATTCCATCTA TGTGTACAAG
  361 GTTCTGAAGC AGGTCCACCC TGACACCGGC ATTTCGTCCA AGGCCATGGG CATCATGAAT
  421 TCGTTTGTGA ACGACATTTT CGAGCGCATC GCAGGTGAGG CTTCCCGCCT GGCGCATTAC
  481 AACAAGCGCT CGACCATCAC CTCCAGGGAG ATCCAGACGG CCGTGCGCCT GCTGCTGCCT
  541 GGGGAGTTGG CCAAGCACGC CGTGTCCGAG GGTACTAAGG CCATCACCAA GTACACCAGC
  601 GCTAAGGATC CAGCCACCAT GGTGAGCAAG GGCGAGGAGG ATAACATGGC CATCATCAAG
  661 GAGTTCATGC GCTTCAAGGT GCACATGGAG GGCTCCGTGA ACGGCCACGA GTTCGAGATC
  721 GAGGGCGAGG GCGAGGGCCG CCCCTACGAN GGCANCCAGA CCGCCAAGCT GAANGTGACC
  781 AAGGGTGGCC CCCTGNCCTT CGCCTGGGAC ATCCTGTCCC CTCAGTTCAT GTACGGCTCC
  841 NAGGCCTACG TNAAGCNNCC CGCCNACATC CCCNANTACT TGAANCTGTC CTTCCCNNNN
  901 GNNTTCNAGN GGGGANCGCG NGATGAACTT CNANGANNGN GGNNTNNNGA NNNNGNNCNN
961 NNNNTNCNTC CNNNNNNNNN GNNNNNNTNN NNCTNNNNNN NNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Kinoshita, N., Diffusible GRAPHIC to visualize morphology of cells after specific cell-cell contact. Sci. Rep. 10 (1): 14437 (2020). PMID 32879377. [link to RRC of NBRP]