Precision gene editing technology and applications in nephrology

Abstract

The expanding field of precision gene editing is empowering researchers to directly modify DNA. Gene editing is made possible using synonymous technologies: a DNA-binding platform to molecularly locate user-selected genomic sequences and an associated biochemical activity that serves as a functional editor. The advent of accessible DNA-targeting molecular systems, such as zinc-finger nucleases, transcription activator-like effectors (TALEs) and CRISPR-Cas9 gene editing systems, has unlocked the ability to target nearly any DNA sequence with nucleotide-level precision. Progress has also been made in harnessing endogenous DNA repair machineries, such as non-homologous end joining, homology-directed repair and microhomology-mediated end joining, to functionally manipulate genetic sequences. As understanding of how DNA damage results in deletions, insertions and modifications increases, the genome becomes more predictably mutable. DNA-binding platforms such as TALEs and CRISPR can also be used to make locus-specific epigenetic changes and to transcriptionally enhance or suppress genes. Although many challenges remain, the application of precision gene editing technology in the field of nephrology has enabled the generation of new animal models of disease as well as advances in the development of novel therapeutic approaches such as gene therapy and xenotransplantation.

Inherited Tubulopathies of the Kidney

“…Yet, for most tubulopathies, treatment has remained essentially unchanged over the past decades and is mainly supportive; for instance, in the form of electrolyte supplements. Fortunately, the advent of gene-editing technologies, such as CRISPR/CAS9 may provide opportunities for the future treatment of tubulopathies (71). In contrast to most other genetic kidney disorders, such as inherited kidney malformations or nephronophthisis, in which global kidney architecture and function is typically irreversibly altered at the time of diagnosis, tubulopathies affect only very specific aspects of kidney function.…”

CAS9 is a genome mutator by directly disrupting DNA-PK dependent DNA repair pathway

“…CAS9 can efficiently target the entire genomic DNA sequence using the Protospacer Adjacent Motif (PAM) domain and a guide RNA (gRNA) (Komor et al, 2017;Murovec et al, 2017). In addition to its extensive use in biomedical research, the clinical application of the CRISPR/CAS9 system in human therapy has been intensively investigated (Barrangou and Doudna, 2016;Dever et al, 2016;Urnov, 2018;WareJoncas et al, 2018;Zhu et al, 2019). Variants of CAS9 have also been developed for distinct biological functions and improved editing fidelity.…”

Tuning up Transcription Factors for Therapy

“…Thus, a TF can be designed to recognize an arbitrary DNA sequence, simply by the permutation of these elements. This combinatorial principle is characteristic of the TALEs, CRISPR-Cas and zinc-finger TFs [3].…”