SHH (sonic hedgehog) is a member of hedgehog family.  SHH is most closely related to Drosophila SHH in terms of sequence and functional similarity. It is instrumental in patterning the early embryo. SHH is implicated as the key inductive signal in patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. SHH activates the transcription of target genes by interacting with its receptor PTCH1 (Patched) to prevent normal inhibition by PTCH1 on the constitutive signaling activity of SMO (smoothened).  SHH is produced as a precursor that is autocatalytically cleaved; the N-terminal portion is soluble and contains the signaling activity while the C-terminal portion is involved in precursor processing with autoproteolysis activity and a cholesterol transferase activity. More interestingly, the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product, restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. The N-product is the active species in both local and long-range signaling, whereas the C-product has no signaling activity.  Cholesterylation is required for N-product targeting to lipid rafts and multimerization. Defects in SHH or in its signalling pathway are a cause of holoprosencephaly (HPE), a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres.

SHH (sonic hedgehog) is a member of hedgehog family.  SHH is most closely related to Drosophila SHH in terms of sequence and functional similarity. It is instrumental in patterning the early embryo. SHH is implicated as the key inductive signal in patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. SHH activates the transcription of target genes by interacting with its receptor PTCH1 (Patched) to prevent normal inhibition by PTCH1 on the constitutive signaling activity of SMO (smoothened).  SHH is produced as a precursor that is autocatalytically cleaved; the N-terminal portion is soluble and contains the signaling activity while the C-terminal portion is involved in precursor processing with autoproteolysis activity and a cholesterol transferase activity. More interestingly, the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product, restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. The N-product is the active species in both local and long-range signaling, whereas the C-product has no signaling activity.  Cholesterylation is required for N-product targeting to lipid rafts and multimerization. Defects in SHH or in its signalling pathway are a cause of holoprosencephaly (HPE), a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres.

 

Product Details

 

Gene Symbol: SHH; TPT; HHG1; HLP3; HPE3; SMMCI; TPTPS; MCOPCB5

 

NCBI Gene ID: 20423

 

Uniprot Entry: Q62226

 

Construct Details: Full length mouse SHH 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: 1314 (bp)

 

Gene Insert Sequence:  

ATGCTGCTGCTGCTGGCCAGATGTTTTCTGGTGATCCTTGCTTCCTCGCTGCTGGTGTGCCCCGGGCTGG

CCTGTGGGCCCGGCAGGGGGTTTGGAAAGAGGCGGCACCCCAAAAAGCTGACCCCTTTAGCCTACAAGCA

GTTTATTCCCAACGTAGCCGAGAAGACCCTAGGGGCCAGCGGCAGATATGAAGGGAAGATCACAAGAAAC

TCCGAACGATTTAAGGAACTCACCCCCAATTACAACCCCGACATCATATTTAAGGATGAGGAAAACACGG

GAGCAGACCGGCTGATGACTCAGAGGTGCAAAGACAAGTTAAATGCCTTGGCCATCTCTGTGATGAACCA

GTGGCCTGGAGTGAAGCTGCGAGTGACCGAGGGCTGGGATGAGGACGGCCATCATTCAGAGGAGTCTCTA

CACTATGAGGGTCGAGCAGTGGACATCACCACGTCCGACCGGGACCGCAGCAAGTACGGCATGCTGGCTC

GCCTGGCTGTGGAAGCAGGTTTCGACTGGGTCTACTATGAATCCAAAGCTCACATCCACTGTTCTGTGAA

AGCAGAGAACTCCGTGGCGGCCAAATCCGGCGGCTGTTTCCCGGGATCCGCCACCGTGCACCTGGAGCAG

GGCGGCACCAAGCTGGTGAAGGACTTACGTCCCGGAGACCGCGTGCTGGCGGCTGACGACCAGGGCCGGC

TGCTGTACAGCGACTTCCTCACCTTCCTGGACCGCGACGAAGGCGCCAAGAAGGTCTTCTACGTGATCGA

GACGCTGGAGCCGCGCGAGCGCCTGCTGCTCACCGCCGCGCACCTGCTCTTCGTGGCGCCGCACAACGAC

TCGGGGCCCACGCCCGGGCCAAGCGCGCTCTTTGCCAGCCGCGTGCGCCCCGGGCAGCGCGTGTACGTGG

TGGCTGAACGCGGCGGGGACCGCCGGCTGCTGCCCGCCGCGGTGCACAGCGTGACGCTGCGAGAGGAGGA

GGCGGGCGCGTACGCGCCGCTCACGGCGCACGGCACCATTCTCATCAACCGGGTGCTCGCCTCGTGCTAC

GCTGTCATCGAGGAGCACAGCTGGGCACACCGGGCCTTCGCGCCTTTCCGCCTGGCGCACGCGCTGCTGG

CCGCGCTGGCACCCGCCCGCACGGACGGCGGGGGCGGGGGCAGCATCCCTGCAGCGCAATCTGCAACGGA

AGCGAGGGGCGCGGAGCCGACTGCGGGCATCCACTGGTACTCGCAGCTGCTCTACCACATTGGCACCTGG

CTGTTGGACAGCGAGACCATGCATCCCTTGGGAATGGCGGTCAAGTCCAGCTGA

 

 

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. Development 120:3339-3353(1994) 

2. Genomics 28:44-51(1995)

3. Nat. Genet. 14:357-360(1996) 

4. Hum. Mol. Genet. 6:1847-1853(1997)

5. J. Biol. Chem. 273:14037-14045(1998)

 

  

 

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.