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
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 of
curing HIV
8. SAMPLE COLLECTION
• Informed consent form
• Blood sample collected from
AF2
• Purification of blood sample to
collect lymphocytes (white
blood cells)
• DNA purification
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 pLNCX2’s
Multiple Cloning Site (MCS).
14. TRANSDUCTION
• RNA is packaged into
retroviral particles
• pLNCX2 does not
contain the structural
genes needed for
particle formation
replication
• Genes are stably
integrated into PT67
• Retroviral particles can
infect target cells
