목차

1. Introduction 1
2. Materials and Methods 4
2.1. Plasmid construction 4
2.2. Prediction of enhancer activity 5
2.3. Cell lines and culture maintenance 5
2.4. Generation and validation of indel mutation cell pools 6
2.5. Generation of EGFP RI pools 6
2.6. Generation of promoter KI monitoring cell line 7
2.7. Generation and validation of ETN KI clone 7
2.8. Generation and validation of EGFP KI clone 8
2.9. Preparation of genomic DNA 8
2.10. Tracking of Indels by DEcomposition (TIDE) analysis 9
2.11. Batch culture 9
2.12. Flow cytometry analysis to measure the levels of EGFP expression 10
2.13. Quantitative real-time polymerase chain reaction 10
2.14. Statistical Analysis 10
3. Results 32
3.1. Identification and characterization of construct integration site 32
3.2. Screening of the IS14 core locus 38
3.3. Assessing the effect of the IS14 sequence 44
3.4. Validation of the IS14 site effects in host cell line 47
3.5. Validation of the IS14 site effects in GS-KO mAb cell line 50
4. Discussion 54
5. References 58