Abstract
CRISPR/Cas (clustered regularly interspaced short palindromic repeats linked to Cas nuclease) technology has revolutionized many aspects of genetic engineering research. Thanks to it, it became possible to study the functions and mechanisms of biology with greater precision, as well as to obtain genetically modified organisms, both prokaryotic and eukaryotic. The changes introduced by the CRISPR/Cas system are based on the repair paths of the single or double strand DNA breaks that cause insertions, deletions, or precise integrations of donor DNA. These changes are crucial for many fields of science, one of which is the use of animals (pigs) as a reservoir of tissues and organs for xenotransplantation into humans. Non-genetically modified animals cannot be used to save human life and health due to acute immunological reactions resulting from the phylogenetic distance of these two species. This review is intended to collect and summarize the advantages as well as achievements of the CRISPR/Cas system in pig-to-human xenotransplantation research. In addition, it demonstrates barriers and limitations that require careful evaluation before attempting to experiment with this technology.
Highlights
The increasing human life expectancy has led to an increase in the number of patients with chronic diseases and severe organ failure
We describe the contribution of this technology to the field of xenotransplantation research and justify its use for this purpose due to the existence of immunological and virological obstacles in pig-to-human xenotransplantation
The authors used kidneys derived from genetically modified pigs with knock-out of porcine GGTA1 gene and knock-in of human CD55 gene, and applied immunosuppression included a transient pan-T cell depletion and an anti-CD154-based regimen combined with daily mycophenolate mofetil (MMF) and solumedrol
Summary
The increasing human life expectancy has led to an increase in the number of patients with chronic diseases and severe organ failure. We describe the CRISPR/Cas system, its use in genetic engineering, its advantages, and its limitations. The CRISPR/Cas system has been described in bacteria and archaea as their acquired ‘immunity’ mechanism against viruses. It works by recognizing and hydrolyzing pathogen genetic material by means of CRISPR RNA (crRNA) associated with CRISPR-associated protein (Cas) nuclease [2,3]. The CRISPR/Cas system has been observed in 40% of bacteria and almost 90% of archaea whose genomes have been sequenced so far [4]. 2021, 22, 3196 bacteria and almost 90% of archaea whose genomes have been sequenced so far [4]. Is used, which is the equivalent of the bacterial chimera: crRNA:tracrRNA (F7i6gure 1)
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