CCR5 Δ32 AS A POTENTIAL GENE

THERAPY FOR AIDS

CONNOR ANDERSON, ALEXANDRA FULTON, MEGAN SHELDON, VICTORIA SOEWARNA & AMANDA WORCESTER

MAKE SURE YOUR VOLUME IS TURNED UP.

THERE ARE VOICEOVERS THROUGHOUT THE PRESENTATION.

PLEASE AND THANK YOU!

HIV ENTRY

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

CCR5 GENE SEQUENCE

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

THE BIG PICTURE• Develop an easier gene therapy technique in hopes of

curing HIV

SAMPLE COLLECTION

• Informed consent form

• Blood sample collected from

AF2

• Purification of blood sample to

collect lymphocytes (white

blood cells)

• DNA purification

CCR5 SEQUENCE

DIGESTIONS• Cut open vector to prepare for

ligation

PLNCX2 MAP

• Designed for retroviral gene delivery and expression

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).

BACTERIAL TRANSFORMATION

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

CULTURE CELLS

• U937 cells possess the CCR5 gene but do not express receptor

FUTURE PLANS

• Test for presence of CCR5 in U937 cells