The length of homology required for gene targeting in embryonic stem cells.
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Implication
Experimental study demonstrates gene targeting efficiency depends on critical homology length in embryonic stem cells, indicating diminishing returns beyond an optimal threshold.
Key Points
- To determine how varying lengths of DNA homology affect the efficiency and frequency of gene targeting in murine embryonic stem cells.
- Measured gene targeting frequencies in murine embryonic stem cells using insertion and replacement vectors with homology lengths ranging from 1.3 to 6.8 kb.
- Evaluated crossover resolution efficiency by comparing replacement vector homology arm lengths of 472 bp against 1.2 kb.
- Gene targeting frequency increased dramatically when sequence homology was extended from 1.3 to 6.8 kb, demonstrating that a critical homology length is required for targeting.
- Extending homology lengths beyond the critical threshold produced progressively smaller increases in gene targeting frequency.
- A homology length of 472 bp was utilized as efficiently as 1.2 kb in the formation and resolution of crossover junctions during gene replacement.
Cite This Study
Hasty et al. (1991) studied this question.