Increasing Gene Editing Efficiency via CRISPR/Cas9- or Cas12a-Mediated Knock-In in Primary Human T Cells

Abstract

T lymphocytes represent a promising target for genome editing. They are primarily modified to recognize and kill tumor cells or to withstand HIV infection. In most studies, T cell genome editing is performed using the CRISPR/Cas technology. Although this technology is easily programmable and widely accessible, its efficiency of T cell genome editing was initially low. Several crucial improvements were made in the components of the CRISPR/Cas technology and their delivery methods, as well as in the culturing conditions of T cells, before a reasonable editing level suitable for clinical applications was achieved. In this review, we summarize and describe the aforementioned parameters that affect human T cell editing efficiency using the CRISPR/Cas technology, with a special focus on gene knock-in.

CRISPR-Cas9 Genome Editing Technology for Zoonotic Disease Control in Indonesia: A Comprehensive Review

“…Due to its ability to target disease vectors such as mosquitoes, ticks, mites, and other arthropods that transmit pathogens to humans and animals and/or vice versa (sylvatic cycle), CRISPR-Cas9 genome editing technology plays a major role in modifying the genetic makeup of these vectors with precision with the aim of reducing or eliminating the ability of these vectors to transmit diseases, with implications for improving the quality of public health [67]. The role and mechanism of targeting disease vectors using CRISPR-Cas9 involves various stages, namely, the identification of target genes that are important for vector competence, reproduction, or survival, especially those that act as pathogen recognition genes, transmission, or vectorspecific traits, such as regulating vector bionomics and reproduction [68,69].…”

“…In addition, advances in immunotherapy models have emerged as a promising approach to combat zoonotic diseases by utilizing the immune system to target and eliminate pathogens [32]. CRISPR technology has enhanced the development of immunotherapies such as monoclonal antibodies, adaptive T-cell therapy, and stem cell therapy to target zoonotic pathogens [68].…”