RESUMEN
Genome editing systems based on site-specific nucleases became very popular for genome editing in modern bioengineering. Human pluripotent stem cells provide a unique platform for genes function study, disease modeling, and drugs testing. Consequently, technology for fast, accurate and well controlled genome manipulation is required. CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRISPR-associated) system could be employed for these purposes. This system is based on site-specific programmable nuclease Cas9. Numerous advantages of the CRISPR/Cas system and its successful application to human stem cells provide wide opportunities for genome therapy and regeneration medicine. In this publication, we describe and compare the main genome editing systems based on site-specific programmable nucleases and discuss opportunities and perspectives of the CRISPR/Cas system for application to pluripotent stem cells.
Asunto(s)
Proteínas Bacterianas/metabolismo , Sistemas CRISPR-Cas , Endonucleasas/metabolismo , Ingeniería Genética/métodos , Genoma Humano , Células Madre Pluripotentes/metabolismo , Proteínas Bacterianas/genética , Roturas del ADN de Doble Cadena , Reparación del ADN por Unión de Extremidades , Bases de Datos Genéticas , Endonucleasas/genética , Recombinación Homóloga , Humanos , Modelos Moleculares , Células Madre Pluripotentes/citología , Medicina Regenerativa/métodos , Streptococcus pyogenes/química , Streptococcus pyogenes/enzimología , Streptococcus pyogenes/genética , Dedos de Zinc/genéticaRESUMEN
Ubiquitin-dependent proteasomal degradation is one of the major pathways of non-lysosomal protein degradation in the cell. The ubiquitination process involves several enzymatic reactions and includes the following enzymes: E1--activating, E2--conjugating, and the third--E3--ligating enzymes. E3 ligases determine the specificity of ubiquitination reaction, i. e. what target protein will be subjected to the covalent modification by ubiquitins. The p53b tumor suppressor protein is one of the most intensively studied over the past several decades. Regulation of its activity is a complex and multi-level process that involves many factors. Ubiquitination is one of the major post-translational modifications of p53, and plays a fundamental role in the control of p53 function, its amount, activity and subcellular localization. This review is focused on p53-specific E3 ubiquitin ligases that are potential targets for cancer therapy using small molecule inhibitors.