FuncLib's Inefficiency in Combinatorial Mutant Generation: Challenges in Gene Synthesis and Gibbs Assembly

Nevertheless, the application of FuncLib in generating multiple combinatorial mutants may prove to be inefficient and uneconomical when subjected to gene synthesis or Gibbs assembly. This is due to the potential adverse effects of upregulation resulting from the mutation, which may further impede the efficacy of FuncLib. The introduction of multiple mutations via FuncLib necessitates the subsequent synthesis of these mutant genes, a process that can be costly and time-consuming. Similarly, employing Gibbs assembly to construct these mutants may also present significant challenges, particularly in terms of scalability and cost-effectiveness. Furthermore, the upregulation associated with these mutations could potentially disrupt the normal cellular processes and lead to undesired phenotypic outcomes, further hindering the efficiency of FuncLib.