![]() Whole exome sequencing was performed to detect the underlying genetic defect in this family. In this study, we determined the genetic cause of split-hand/foot malformation in a Chinese pedigree.įor this study, we have recruited a Chinese family and collected samples from affected and normal individuals of the family (three affected and two normal). Variants associated with the TP63 gene are known to cause multiple disorders. ![]() Tumor protein p63 is an important transcription factor regulating epithelial morphogenesis. OrthoRep enables a new paradigm of routine, high-throughput evolution of biomolecular and cellular function. We uncovered a more complex fitness landscape than previously realized, including common adaptive trajectories constrained by epistasis, rare outcomes that avoid a frequent early adaptive mutation, and a suboptimal fitness peak that occasionally traps evolving populations. Using OrthoRep, we evolved drug-resistant malarial dihydrofolate reductases (DHFRs) in 90 independent replicates. User-defined genes in OrthoRep continuously and rapidly evolve through serial passaging, a highly straightforward and scalable process. We report OrthoRep, an orthogonal DNA polymerase-plasmid pair in yeast that stably mutates ∼100,000-fold faster than the host genome in vivo, exceeding the error threshold of genomic replication that causes single-generation extinction. However, experimental strategies for directed evolution are notoriously labor intensive and low throughput, limiting access to demanding functions, multiple functions in parallel, and the study of molecular evolution in replicate. ![]() Directed evolution is a powerful approach for engineering biomolecules and understanding adaptation. ![]()
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