Mouse Models of Prostate Cancer

Name: Vicki Javier

Mentor: Todd Hricik

Institution: MSKCC

• Prostate Cancer: disease marked by abnormal growth of tumor cells in the prostate gland

• Prostate Cancer is detected by– Digital Rectal Exam– Biopsy– PSA serum reading

Prostate Cancer

• PSA is a biomarker for CaP

• Our Question: Does PSA play a role in CaP?-Mouse studies

PSA Serum Screen

PSA mouse study

• Mice don’t have PSA

• We already put WT PSA gene into mice

• We now need a mutant form of PSA gene to put in mice so that we can compare whether the mut mouse will develop CaP in a different way than the WT PSA mouse

• Mutate the PSA gene (in a plasmid) nucleotide sequence so that a Serine amino acid is changed to an Alanine in the PSA protein (gene product)

• Prepare the mutated PSA gene for injection into the mouse

Project Goal

Promoter WT PSA Ser 189 SV40 Poly A

NotIKpnI

EcoRI

EcoRI

Promoter Mut PSA Ser 189 to Ala SV40 Poly A

NotIKpnI

EcoRI

EcoRI

SDM

Site Directed Mutagenesis

TCT = Serine

GCT = Alanine

Tools

Restriction Digestion of DNA

Direction of DNA travel

6000 bp5000 bp

250 bp

References

• Hricik, Todd

• http://www.web-books.com/MoBio/Free/Ch9G.htm

• http://en.wikipedia.org/wiki/Site-directed_mutagenesis

• http://www.ehealthmd.com/library/prostateremoval/PR_glossary.html

Acknowledgements

• Todd Hricik

• Dr. Sat Bhattacharya

• Sven

• Ross

• Neil