1. 1. CCR5 32 AS APOTENTIAL GENE THERAPY FOR AIDS CONNOR ANDERSON, ALEXANDRA FULTON, MEGAN SHELDON, VICTORIASOEWARNA & AMANDA WORCESTER
2. 2. MAKE SURE YOUR VOLUME ISTURNED UP.THERE ARE VOICEOVERSTHROUGHOUT THEPRESENTATION.PLEASE AND THANK YOU!
3. 3. HIV ENTRY
4. 4. DELTA 32 Current deletion mutation that results in a frame shift 32 base pairs are absent from the gene sequence THE CAT AND DOG RAN FOR FUN A THE CTA NDD OGR ANF ORF UN
5. 5. CCR5 GENE SEQUENCE
6. 6. TIMOTHY RAY BROWN Had HIV and Leukemia Leukemia was eradicated from the body HIV status was reversed Genetically altered genes into a vector Vector to cell line Gene therapy and treatment
7. 7. THE BIG PICTURE Develop an easier gene therapy technique in hopes ofcuring HIV
8. 8. SAMPLE COLLECTION Informed consent form Blood sample collected fromAF2 Purification of blood sample tocollect lymphocytes (whiteblood cells) DNA purification
9. 9. CCR5 SEQUENCE
10. 10. DIGESTIONS Cut open vector to prepare forligation
11. 11. PLNCX2 MAP Designed for retroviral gene delivery and expression
12. 12. LIGATION Ligated oligonucleotides onto AF2 Converted BamHI into HINDIII site, and Xba into NOTI site. Made AF2 compatible for ligation into pLNCX2sMultiple Cloning Site (MCS).
13. 13. BACTERIAL TRANSFORMATION
14. 14. TRANSDUCTION RNA is packaged intoretroviral particles pLNCX2 does notcontain the structuralgenes needed forparticle formationreplication Genes are stablyintegrated into PT67 Retroviral particles caninfect target cells
15. 15. CULTURE CELLS U937 cells possess the CCR5 gene but do notexpress receptor
16. 16. FUTURE PLANS Test for presence of CCR5 in U937 cells