MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems

@article{Sakuma2016MMEJassistedGK,
  title={MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems},
  author={Tetsushi Sakuma and Shota Nakade and Yuto Sakane and Ken-Ichi T Suzuki and Takashi Yamamoto},
  journal={Nature Protocols},
  year={2016},
  volume={11},
  pages={118-133}
}
Programmable nucleases enable engineering of the genome by utilizing endogenous DNA double-strand break (DSB) repair pathways. Although homologous recombination (HR)-mediated gene knock-in is well established, it cannot necessarily be applied in every cell type and organism because of variable HR frequencies. We recently reported an alternative method of gene knock-in, named the PITCh (Precise Integration into Target Chromosome) system, assisted by microhomology-mediated end-joining (MMEJ… CONTINUE READING