1. CCR5 32 AS APOTENTIAL GENE THERAPY FOR AIDS CONNOR ANDERSON,
ALEXANDRA FULTON, MEGAN SHELDON, VICTORIASOEWARNA & AMANDA
WORCESTER
2. MAKE SURE YOUR VOLUME ISTURNED UP.THERE ARE
VOICEOVERSTHROUGHOUT THEPRESENTATION.PLEASE AND THANK YOU!
3. HIV ENTRY
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. CCR5 GENE SEQUENCE
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. THE BIG PICTURE Develop an easier gene therapy technique in
hopes ofcuring HIV
8. SAMPLE COLLECTION Informed consent form Blood sample
collected fromAF2 Purification of blood sample tocollect
lymphocytes (whiteblood cells) DNA purification
9. CCR5 SEQUENCE
10. DIGESTIONS Cut open vector to prepare forligation
11. PLNCX2 MAP Designed for retroviral gene delivery and
expression
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. BACTERIAL TRANSFORMATION
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. CULTURE CELLS U937 cells possess the CCR5 gene but do
notexpress receptor
16. FUTURE PLANS Test for presence of CCR5 in U937 cells