bioRxivBiochemistry
Addressing the dNTP bottleneck restricting prime editing activity
K. Ponnienselvan, P. Liu, T. Nyalile, S. Oikemus, A. T. Joynt, K. Kelly, D. Guo, Z. Chen, J. M. Lee, C. A. Schiffer, C. P. Emerson, N. D. Lawson, J. K. Watts, E. J. Sontheimer, J. Luban, S. A. Wolfe
Abstract
Prime editing efficiency is modest in cells that are quiescent or slowly proliferating where intracellular dNTP levels are tightly regulated. MMLV-reverse transcriptase - the prime editor polymerase subunit - requires high intracellular dNTPs levels for efficient polymerization. We report that prime editing efficiency in primary cells and in vivo is increased by mutations that enhance the enzymatic properties of MMLV-reverse transcriptase and can be further complemented by targeting SAMHD1 for degradation.
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