A survey of barcoding technology, its use in cancer biology, and my work in optimizing the lentiviral supernatant concentration.

1. Department of Oncology
Gregory Schwing
Gawad Lab
Mentor: Dr Gawad, MD, PhD
St Jude
8/7/15

3. American Society of Hematology et al 2012
3000 children are diagnosed
with ALL each year

4. • Approximately 600 cases of TELAML1
every year
• TELAML1 occurs in utero
• TELAML1 causes a halt in the
development of B-cell progenitor cells
• 60,000 children are born with the
TELAML1 mutation each year, a state
we call Pre-Leukemic

5. mRNA editing
Retrovirus infection
2 functions:
(Harjes, Gross et al. 2009)

6. Gawad et al 2014

7. • Unique heritable genetic tag
• Frequency of barcode measures the fitness of a parent cell
GACTCTCTGAG
GACTCTCTGAG
GACTCTCTGAG
http://www.dreamstime.com/

8. (Levy, Blundell et al. 2015)

10. GAG/POL REV/TAT ENV
Human Embryonic Kidney Cell

12. Dilution Factor
%GFPExpression

13. • The barcoded vector was successfully produced in
Lentivirus
• We determined the optimal concentration for viral
transduction is the undiluted supernatant

14. • Analyze barcode diversity
• Transduce the barcoded virus into the hematopoetic stem
cells, graft the HSC into mice, and monitor the barcodes.
• Transduce APOBEC into the HSC coexpressed with ETV6-
RUNX1 to test our model

15. Acknowledgements