RTN4 is a two-pass type II transmembrane protein belonging to the reticulon family. Reticulons are associated with the endoplasmic reticulum, and are involved in neuroendocrine secretion or in membrane trafficking in neuroendocrine cells.  RTN4 is a potent neurite outgrowth inhibitor which may also help block the regeneration of the central nervous system in higher vertebrates. Multiple isoforms of RTN4 have been described.  Isoform 1 is specifically expressed in brain and testis and weakly in heart and skeletal muscle. Isoform 2 is widely expressed except for the liver. Isoform 3 is expressed in brain, skeletal muscle and adipocytes. Isoform 4 is testis-specific. RTN4 isoform 1 binds to RTN4R. Isoform 2 binds to NGBR and RTN3. Isoform 2 and isoform 3 interact with BACE1 and BACE2. Isoform 2 reduces the anti-apoptotic activity of Bcl-xl and Bcl-2. Isoform 2 and isoform 3 inhibit BACE1 activity and amyloid precursor protein processing. Three regions, residues 59-172, 544-725 and the loop 66 amino acids, between the two transmembrane domains, known as Nogo-66 loop, appear to be responsible for the inhibitory effect on neurite outgrowth and the spreading of neurons. This Nogo-66 loop, mediates also the binding of RTN4 to its receptor. RTN4 acts as a negative regulator of axon-axon adhesion and growth, and as a facilitator of neurite branching.  RTN4 down-regulation of growth, stabilization of wiring and restriction of plasticity in the adult CNS and mediates the radial migration of cortical neurons via an RTN4R-LINGO1 containing receptor complex. 

RTN4 is a two-pass type II transmembrane protein belonging to the reticulon family. Reticulons are associated with the endoplasmic reticulum, and are involved in neuroendocrine secretion or in membrane trafficking in neuroendocrine cells.  RTN4 is a potent neurite outgrowth inhibitor which may also help block the regeneration of the central nervous system in higher vertebrates. Multiple isoforms of RTN4 have been described.  Isoform 1 is specifically expressed in brain and testis and weakly in heart and skeletal muscle. Isoform 2 is widely expressed except for the liver. Isoform 3 is expressed in brain, skeletal muscle and adipocytes. Isoform 4 is testis-specific. RTN4 isoform 1 binds to RTN4R. Isoform 2 binds to NGBR and RTN3. Isoform 2 and isoform 3 interact with BACE1 and BACE2. Isoform 2 reduces the anti-apoptotic activity of Bcl-xl and Bcl-2. Isoform 2 and isoform 3 inhibit BACE1 activity and amyloid precursor protein processing. Three regions, residues 59-172, 544-725 and the loop 66 amino acids, between the two transmembrane domains, known as Nogo-66 loop, appear to be responsible for the inhibitory effect on neurite outgrowth and the spreading of neurons. This Nogo-66 loop, mediates also the binding of RTN4 to its receptor. RTN4 acts as a negative regulator of axon-axon adhesion and growth, and as a facilitator of neurite branching.  RTN4 down-regulation of growth, stabilization of wiring and restriction of plasticity in the adult CNS and mediates the radial migration of cortical neurons via an RTN4R-LINGO1 containing receptor complex. 

 

Product Details

 

Gene Symbol: RTN4; ASY; NSP; NOGO; NOGOC; RTN-X; NOGO-A; NSP-CL; Nogo-B; Nogo-C; RTN4-A; RTN4-C; RTN4-B1; RTN4-B2; NI220/250; Nbla00271; Nbla10545

 

NCBI Gene ID: 57142

 

Uniprot Entry: Q9NQC3

 

Construct Details: Full length human RTN4 gene is subcloned into the Lentiviral expression vector pLTC with an upstream CMV promoter and with or without a selection marker, which can be used for both transient and stable expression in mammalian cells.  It can be co-transfected with the LentiPAK DNA mix (SKU# LP-001) into HEK293 cells to produce high titer lentiviral particles. Multiple choices of a stable antibiotic selection marker are available.

 

Vector Type: pLTC or pLTC-IRES-Marker (lentiviral expression vector containing a heterologous CMV promoter, see the vector map above)

 

Formulation: Supplied with 10 μg of endotoxin-free plasmid DNA at 200 ng/μl in sterile TE buffer (pH8.0)

 

Gene Insert Size: 3579 (bp)

 

Gene Insert Sequence:  

ATGGAAGACCTGGACCAGTCTCCTCTGGTCTCGTCCTCGGACAGCCCACCCCGGCCGCAGCCCGCGTTCA

AGTACCAGTTCGTGAGGGAGCCCGAGGACGAGGAGGAAGAAGAGGAGGAGGAAGAGGAGGACGAGGACGA

AGACCTGGAGGAGCTGGAGGTGCTGGAGAGGAAGCCCGCCGCCGGGCTGTCCGCGGCCCCAGTGCCCACC

GCCCCTGCCGCCGGCGCGCCCCTGATGGACTTCGGAAATGACTTCGTGCCGCCGGCGCCCCGGGGACCCC

TGCCGGCCGCTCCCCCCGTCGCCCCGGAGCGGCAGCCGTCTTGGGACCCGAGCCCGGTGTCGTCGACCGT

GCCCGCGCCATCCCCGCTGTCTGCTGCCGCAGTCTCGCCCTCCAAGCTCCCTGAGGACGACGAGCCTCCG

GCCCGGCCTCCCCCTCCTCCCCCGGCCAGCGTGAGCCCCCAGGCAGAGCCCGTGTGGACCCCGCCAGCCC

CGGCTCCCGCCGCGCCCCCCTCCACCCCGGCCGCGCCCAAGCGCAGGGGCTCCTCGGGCTCAGTGGATGA

GACCCTTTTTGCTCTTCCTGCTGCATCTGAGCCTGTGATACGCTCCTCTGCAGAAAATATGGACTTGAAG

GAGCAGCCAGGTAACACTATTTCGGCTGGTCAAGAGGATTTCCCATCTGTCCTGCTTGAAACTGCTGCTT

CTCTTCCTTCTCTGTCTCCTCTCTCAGCCGCTTCTTTCAAAGAACATGAATACCTTGGTAATTTGTCAAC

AGTATTACCCACTGAAGGAACACTTCAAGAAAATGTCAGTGAAGCTTCTAAAGAGGTCTCAGAGAAGGCA

AAAACTCTACTCATAGATAGAGATTTAACAGAGTTTTCAGAATTAGAATACTCAGAAATGGGATCATCGT

TCAGTGTCTCTCCAAAAGCAGAATCTGCCGTAATAGTAGCAAATCCTAGGGAAGAAATAATCGTGAAAAA

TAAAGATGAAGAAGAGAAGTTAGTTAGTAATAACATCCTTCATAATCAACAAGAGTTACCTACAGCTCTT

ACTAAATTGGTTAAAGAGGATGAAGTTGTGTCTTCAGAAAAAGCAAAAGACAGTTTTAATGAAAAGAGAG

TTGCAGTGGAAGCTCCTATGAGGGAGGAATATGCAGACTTCAAACCATTTGAGCGAGTATGGGAAGTGAA

AGATAGTAAGGAAGATAGTGATATGTTGGCTGCTGGAGGTAAAATCGAGAGCAACTTGGAAAGTAAAGTG

GATAAAAAATGTTTTGCAGATAGCCTTGAGCAAACTAATCACGAAAAAGATAGTGAGAGTAGTAATGATG

ATACTTCTTTCCCCAGTACGCCAGAAGGTATAAAGGATCGTTCAGGAGCATATATCACATGTGCTCCCTT

TAACCCAGCAGCAACTGAGAGCATTGCAACAAACATTTTTCCTTTGTTAGGAGATCCTACTTCAGAAAAT

AAGACCGATGAAAAAAAAATAGAAGAAAAGAAGGCCCAAATAGTAACAGAGAAGAATACTAGCACCAAAA

CATCAAACCCTTTTCTTGTAGCAGCACAGGATTCTGAGACAGATTATGTCACAACAGATAATTTAACAAA

GGTGACTGAGGAAGTCGTGGCAAACATGCCTGAAGGCCTGACTCCAGATTTAGTACAGGAAGCATGTGAA

AGTGAATTGAATGAAGTTACTGGTACAAAGATTGCTTATGAAACAAAAATGGACTTGGTTCAAACATCAG

AAGTTATGCAAGAGTCACTCTATCCTGCAGCACAGCTTTGCCCATCATTTGAAGAGTCAGAAGCTACTCC

TTCACCAGTTTTGCCTGACATTGTTATGGAAGCACCATTGAATTCTGCAGTTCCTAGTGCTGGTGCTTCC

GTGATACAGCCCAGCTCATCACCATTAGAAGCTTCTTCAGTTAATTATGAAAGCATAAAACATGAGCCTG

AAAACCCCCCACCATATGAAGAGGCCATGAGTGTATCACTAAAAAAAGTATCAGGAATAAAGGAAGAAAT

TAAAGAGCCTGAAAATATTAATGCAGCTCTTCAAGAAACAGAAGCTCCTTATATATCTATTGCATGTGAT

TTAATTAAAGAAACAAAGCTTTCTGCTGAACCAGCTCCGGATTTCTCTGATTATTCAGAAATGGCAAAAG

TTGAACAGCCAGTGCCTGATCATTCTGAGCTAGTTGAAGATTCCTCACCTGATTCTGAACCAGTTGACTT

ATTTAGTGATGATTCAATACCTGACGTTCCACAAAAACAAGATGAAACTGTGATGCTTGTGAAAGAAAGT

CTCACTGAGACTTCATTTGAGTCAATGATAGAATATGAAAATAAGGAAAAACTCAGTGCTTTGCCACCTG

AGGGAGGAAAGCCATATTTGGAATCTTTTAAGCTCAGTTTAGATAACACAAAAGATACCCTGTTACCTGA

TGAAGTTTCAACATTGAGCAAAAAGGAGAAAATTCCTTTGCAGATGGAGGAGCTCAGTACTGCAGTTTAT

TCAAATGATGACTTATTTATTTCTAAGGAAGCACAGATAAGAGAAACTGAAACGTTTTCAGATTCATCTC

CAATTGAAATTATAGATGAGTTCCCTACATTGATCAGTTCTAAAACTGATTCATTTTCTAAATTAGCCAG

GGAATATACTGACCTAGAAGTATCCCACAAAAGTGAAATTGCTAATGCCCCGGATGGAGCTGGGTCATTG

CCTTGCACAGAATTGCCCCATGACCTTTCTTTGAAGAACATACAACCCAAAGTTGAAGAGAAAATCAGTT

TCTCAGATGACTTTTCTAAAAATGGGTCTGCTACATCAAAGGTGCTCTTATTGCCTCCAGATGTTTCTGC

TTTGGCCACTCAAGCAGAGATAGAGAGCATAGTTAAACCCAAAGTTCTTGTGAAAGAAGCTGAGAAAAAA

CTTCCTTCCGATACAGAAAAAGAGGACAGATCACCATCTGCTATATTTTCAGCAGAGCTGAGTAAAACTT

CAGTTGTTGACCTCCTGTACTGGAGAGACATTAAGAAGACTGGAGTGGTGTTTGGTGCCAGCCTATTCCT

GCTGCTTTCATTGACAGTATTCAGCATTGTGAGCGTAACAGCCTACATTGCCTTGGCCCTGCTCTCTGTG

ACCATCAGCTTTAGGATATACAAGGGTGTGATCCAAGCTATCCAGAAATCAGATGAAGGCCACCCATTCA

GGGCATATCTGGAATCTGAAGTTGCTATATCTGAGGAGTTGGTTCAGAAGTACAGTAATTCTGCTCTTGG

TCATGTGAACTGCACGATAAAGGAACTCAGGCGCCTCTTCTTAGTTGATGATTTAGTTGATTCTCTGAAG

TTTGCAGTGTTGATGTGGGTATTTACCTATGTTGGTGCCTTGTTTAATGGTCTGACACTACTGATTTTGG

CTCTCATTTCACTCTTCAGTGTTCCTGTTATTTATGAACGGCATCAGGCACAGATAGATCATTATCTAGG

ACTTGCAAATAAGAATGTTAAAGATGCTATGGCTAAAATCCAAGCAAAAATCCCTGGATTGAAGCGCAAA

GCTGAATGA

 

 

Primers: Gene insert coding sequence can be confirmed by following primers:

(a) forward (or 5'-end) primer: 5’-CAAATGGGCGGTAGGCGTG-3’;

(b) reverse (or 3'-end) primer: 5’-CCAGGTTTCCGGGCCCTCAC-3’

 

 

Note: The sequence may vary due to the naturally occurring variants or mutations, such as single nucleotide polymorphism, or the artifact during PCR amplification.  High fidelity PCR systems should always be used. 

 

 

  

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

 

Storage

 

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 6 months. 

 

 

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund. 

 

References

 

1. Nature 409:341 (2001) 

2. Nature 417:547 (2002)

3. Nature 417:941 (2002)

4. Science 297:1190 (2002)

5. J. Neurosci. Res. 67:559 2002) 

6. Nat. Neurosci. 7:221 (2004)

 

  

 

Additional supporting documents, including product datasheet, COA and MSDS are available upon request.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.