1. KLF1 Effects on γ-Globin Regulation
Ashria Arora

2. β– and γ – Globin Genes Beta globin expressed in adulthood
Gamma globin expressed fetal erythroblasts
Transcription factors and repressor involved
Mutation in the upstream promoter can cause:
-beta thalasemia
~production of low to none beta globin
~production and expression of γ – globin to compensate for beta

3. KLF1 Krüppel-like transcription factors
-Krüppel is a gap gene, encodes zinc fingers
~Cys/His zinc fingers
-Family of 17 factors
-Previous studies proven KLF1 regulates transcription of beta globin
~KLF1 null can cause beta thalasemia- patient with null

4. Research Question
Is the overexpression of KLF1 a positive or negative regulator γ-globin genes?

5. Experiment
Human fetal erythroid cell line along with zebrafish fetal cell line
-control- zebrafish, human adult cell with no plasmid construct
Plasmid constructs
Luciferase assay

6. Methods Plasmid constructs -KLF1, LUC, and γCACCCLuc will be generated
-KLF1 replicated inside human cell line to produce the overexpression of KLF1 (recobinant DNA)
-Generating expression constructs, luciferase assay
Luciferase assay- firefly enzyme
Emission of light is measured to determine expression of γ-globin genes, CAACCLuc allows
More light, mean transcription of γ-globin

7. Possible Results
Zebrafish represents the expression w/o KLF1, human with normal levels of KLF1, and human with the overexpression
Possible outcomes:
Light emitted to be similar to control (human cell line, normal levels), overexprssion has no effect
Light emitted to be less than control, therefore overexpression negatively regulates γ-globin
Light emitted more than control levels, overexpression positively regulates