Human TIGIT/VSIG9/VSTM3/WUCAM Gene, Full-length cDNA Clone in Lentivector, Endotoxin-free DNA

Description

TIGIT (T cell immunoreceptor with immunoglobulin and ITIM domains), also known as VSTM3 (V-set and transmembrane domain-containing protein 3) and VSIG9 (V-set and immunoglobulin domain-containing protein 9), is a single pass type I transmembrane protein belonging to the CD28 family within the Ig superfamily (IgSF).  TIGIT contains a 115 aa extracellular region with a V-type Ig­like (IgV) domain, a transmembrane (TM) domain and a 79 aa cytoplasmic region with an immunoreceptor tyrosine-based inhibitory motif (ITIM). TIGIT is expressed on NK cells and regulatory, memory, activated T cells including follicular helper T cells within secondary lymphoid organs. TIGIT binds to CD155/PVR/Necl5 with high affinity and CD112/PVRL2/Nectin2 with low affinity. Through CD155 and CD112, TIGIT also interacts with other IgSF members such as CD226/DNAM1 and CD96/Tactile. TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells (DCs). The binding of PVR to TIGIT on human DCs induces the production of IL-10 and inhibits the production of IL-12. Ligation of TIGIT on T cells down-regulates TCR­mediated activation and T proliferation, while TIGIT ligation on NK cells blocks NK cell cytotoxicity directly through its ITIM. Therefore TIGIT inhibits the NK-mediated killing of tumor cells and protects normal cells from NK-mediated cytotoxicity. Mice deficient in TIGIT show increased T cell responses and susceptibility to autoimmune challenges. Soluble TIGIT is able to compete with CD226 for CD155 binding and attenuate T cell responses. 

 

 

Gene Symbol: TIGIT; VSIG9; VSTM3; WUCAM

 

NCBI Gene ID: 201633

 

Uniprot Entry: Q495A1

 

Construct Details: Full length human TIGIT gene (encoding UniProt #Q495A1) 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: 735 (bp)

 

Gene Insert Sequence:  

ATGCGCTGGTGTCTCCTCCTGATCTGGGCCCAGGGGCTGAGGCAGGCTCCCCTCGCCTCAGGAATGATGACAGGCACAAT

AGAAACAACGGGGAACATTTCTGCAGAGAAAGGTGGCTCTATCATCTTACAATGTCACCTCTCCTCCACCACGGCACAAG

TGACCCAGGTCAACTGGGAGCAGCAGGACCAGCTTCTGGCCATTTGTAATGCTGACTTGGGGTGGCACATCTCCCCATCC

TTCAAGGATCGAGTGGCCCCAGGTCCCGGCCTGGGCCTCACCCTCCAGTCGCTGACCGTGAACGATACAGGGGAGTACTT

CTGCATCTATCACACCTACCCTGATGGGACGTACACTGGGAGAATCTTCCTGGAGGTCCTAGAAAGCTCAGTGGCTGAGC

ACGGTGCCAGGTTCCAGATTCCATTGCTTGGAGCCATGGCCGCGACGCTGGTGGTCATCTGCACAGCAGTCATCGTGGTG

GTCGCGTTGACTAGAAAGAAGAAAGCCCTCAGAATCCATTCTGTGGAAGGTGACCTCAGGAGAAAATCAGCTGGACAGGA

GGAATGGAGCCCCAGTGCTCCCTCACCCCCAGGAAGCTGTGTCCAGGCAGAAGCTGCACCTGCTGGGCTCTGTGGAGAGC

AGCGGGGAGAGGACTGTGCCGAGCTGCATGACTACTTCAATGTCCTGAGTTACAGAAGCCTGGGTAACTGCAGCTTCTTC

ACAGAGACTGGTTAG

 

 

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.

 

 

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.

 

 

1. Eur. J. Immunol. 39:695 (2009).

2. Nat. Immunol. 10:48 (2009). 

3. Proc. Natl. Acad. Sci.106:17858 (2009). 

4. J. Immunol. 186:1338 (2011).

5. Eur. J. Immunol. 41:902 (2011).

 

 

 

 

Product datasheet (pdf) can be downloaded here: LTP0963-PDS.pdf

 

 

 

Additional supporting documents, including 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.