IGEM:Harvard/2010/Team Allergy

Description

We are designing shRNA constructs against common allergens in fruits and vegetables.

Bibliography

1. Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, Green AG, and Waterhouse PM. Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 2001 Sep;27(6):581-90. DOI:10.1046/j.1365-313x.2001.01105.x |
2. Dodo HW, Konan KN, Chen FC, Egnin M, and Viquez OM. Alleviating peanut allergy using genetic engineering: the silencing of the immunodominant allergen Ara h 2 leads to its significant reduction and a decrease in peanut allergenicity. Plant Biotechnol J. 2008 Feb;6(2):135-45. DOI:10.1111/j.1467-7652.2007.00292.x |
3. Chardin H, Mayer C, Sénéchal H, Wal JM, Poncet P, Desvaux FX, and Peltre G. Lipid transfer protein 1 is a possible allergen in Arabidopsis thaliana. Int Arch Allergy Immunol. 2003 Jun;131(2):85-90. DOI:10.1159/000070923 |
4. San Miguel-Moncín M, Krail M, Scheurer S, Enrique E, Alonso R, Conti A, Cisteró-Bahíma A, and Vieths S. Lettuce anaphylaxis: identification of a lipid transfer protein as the major allergen. Allergy. 2003 Jun;58(6):511-7. DOI:10.1034/j.1398-9995.2003.00097.x |
5. Salcedo G, Sanchez-Monge R, Diaz-Perales A, Garcia-Casado G, and Barber D. Plant non-specific lipid transfer proteins as food and pollen allergens. Clin Exp Allergy. 2004 Sep;34(9):1336-41. DOI:10.1111/j.1365-2222.2004.02018.x |
6. Gajhede M, Osmark P, Poulsen FM, Ipsen H, Larsen JN, Joost van Neerven RJ, Schou C, Løwenstein H, and Spangfort MD. X-ray and NMR structure of Bet v 1, the origin of birch pollen allergy. Nat Struct Biol. 1996 Dec;3(12):1040-5. DOI:10.1038/nsb1296-1040 |
7. Zuidmeer L, Salentijn E, Rivas MF, Mancebo EG, Asero R, Matos CI, Pelgrom KT, Gilissen LJ, and van Ree R. The role of profilin and lipid transfer protein in strawberry allergy in the Mediterranean area. Clin Exp Allergy. 2006 May;36(5):666-75. DOI:10.1111/j.1365-2222.2006.02453.x |
8. Karlsson AL, Alm R, Ekstrand B, Fjelkner-Modig S, Schiött A, Bengtsson U, Björk L, Hjernø K, Roepstorff P, and Emanuelsson CS. Bet v 1 homologues in strawberry identified as IgE-binding proteins and presumptive allergens. Allergy. 2004 Dec;59(12):1277-84. DOI:10.1111/j.1398-9995.2004.00585.x |
9. Musidlowska-Persson A, Alm R, and Emanuelsson C. Cloning and sequencing of the Bet v 1-homologous allergen Fra a 1 in strawberry (Fragaria ananassa) shows the presence of an intron and little variability in amino acid sequence. Mol Immunol. 2007 Feb;44(6):1245-52. DOI:10.1016/j.molimm.2006.06.004 |
10. Breiteneder H and Ebner C. Molecular and biochemical classification of plant-derived food allergens. J Allergy Clin Immunol. 2000 Jul;106(1 Pt 1):27-36. DOI:10.1067/mai.2000.106929 |
11. Singh MB and Bhalla PL. Genetic engineering for removing food allergens from plants. Trends Plant Sci. 2008 Jun;13(6):257-60. DOI:10.1016/j.tplants.2008.04.004 |
12. Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, Green AG, and Waterhouse PM. Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 2001 Sep;27(6):581-90. DOI:10.1046/j.1365-313x.2001.01105.x |
13. Dodo HW, Konan KN, Chen FC, Egnin M, and Viquez OM. Alleviating peanut allergy using genetic engineering: the silencing of the immunodominant allergen Ara h 2 leads to its significant reduction and a decrease in peanut allergenicity. Plant Biotechnol J. 2008 Feb;6(2):135-45. DOI:10.1111/j.1467-7652.2007.00292.x |
14. Le LQ, Mahler V, Lorenz Y, Scheurer S, Biemelt S, Vieths S, and Sonnewald U. Reduced allergenicity of tomato fruits harvested from Lyc e 1-silenced transgenic tomato plants. J Allergy Clin Immunol. 2006 Nov;118(5):1176-83. DOI:10.1016/j.jaci.2006.06.031 |

Primers and Sequences

name	GenBank Accession	forward sense primer	reverse sense primer	forward antisense primer	reverse antisense primer
pdk intron	AJ311872	CCTTGAATTCGCGGCCGCATCTAGA tcttttttccttttagtata	AAGGCTGCAGCGGCCGCTACTAGT CAATCCAAATGTAAGATCAA
A. thaliana LTP1	ATHSEQB	CCTTGAATTCGCGGCCGCATCTAGA ATGGCTGGAGTGATGAAGTT	AAGGCTGCAGCGGCCGCTACTAGT CAGCTGCACGGCCAGCGTTG	AAGGCTGCAGCGGCCGCTACTAGT ATGGCTGGAGTGATGAAGTT	CCTTGAATTCGCGGCCGCATCTAGA CAGCTGCACGGCCAGCGTTG
Fra a 1A	DQ385511	CCTTGAATTCGCGGCCGCATCTAGA ATGGGTGTTTACACTTATGA	AAGGCTGCAGCGGCCGCTACTAGT GTAGTCGATCTTCTCGATGT	AAGGCTGCAGCGGCCGCTACTAGT ATGGGTGTTTACACTTATGA	CCTTGAATTCGCGGCCGCATCTAGA GTAGTCGATCTTCTCGATGT
L. sativa LTP 1.1	EF101532.1	CCTTGAATTCGCGGCCGCATCTAGA ATGGCAAGGATGGCAACGAT	AAGGCTGCAGCGGCCGCTACTAGT TGACACCACACTTGCCGGGG	AAGGCTGCAGCGGCCGCTACTAGT ATGGCAAGGATGGCAACGAT	CCTTGAATTCGCGGCCGCATCTAGA TGACACCACACTTGCCGGGG
L. sativa LTP 1.2	EF101531.1	CCTTGAATTCGCGGCCGCATCTAGA GTGACATGTGGTCAGGTGGT	AAGGCTGCAGCGGCCGCTACTAGT CTGGATCCTATTGCAGTCGG	AAGGCTGCAGCGGCCGCTACTAGT GTGACATGTGGTCAGGTGGT	CCTTGAATTCGCGGCCGCATCTAGA CTGGATCCTATTGCAGTCGG
		CCTTGAATTCGCGGCCGCATCTAGA	AAGGCTGCAGCGGCCGCTACTAGT	AAGGCTGCAGCGGCCGCTACTAGT	CCTTGAATTCGCGGCCGCATCTAGA
Lyc E 1.01 editing	AJ417553	CCTTGAATTCGCGGCCGCATCTAGA	AAGGCTGCAGCGGCCGCTACTAGT	AAGGCTGCAGCGGCCGCTACTAGT	CCTTGAATTCGCGGCCGCATCTAGA