The discovery of CRISPR/Cas9 brought a hope for having an efficient, reliable, and readily available tool for genome editing. CRISPR/Cas9 is certainly easy to use, while its efficiency and reliability remain the focus of studies. The review describes the general principles of the organization and function of Cas nucleases and a number of important issues to be considered while planning genome editing experiments with CRISPR/Cas9. The issues include evaluation of the efficiency and specificity for Cas9, sgRNA selection, Cas9 variants designed artificially, and use of homologous recombination and nonhomologous end joining in DNA editing.
Bacterial Proteins/genetics , CRISPR-Cas Systems , Clustered Regularly Interspaced Short Palindromic Repeats , Endonucleases/genetics , Gene Editing/methods , Genome , RNA, Guide, Kinetoplastida/genetics , Animals , Bacterial Proteins/metabolism , Base Pairing , Base Sequence , CRISPR-Associated Protein 9 , DNA End-Joining Repair , Endonucleases/metabolism , Humans , Protein Engineering , RNA, Guide, Kinetoplastida/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Recombinational DNA Repair
Depending on the type of autonomous regulation, differences in basic levels of interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) were revealed under conditions of hyperthermia in healthy subjects aged 19-21. A parasympathetic type of autonomous regulation corresponded to higher initial levels of proinflammatory cytokinesis, whereas a dominating sympathetic type corresponded to lower levels of the IL-1 beta and TNF alpha. The subjects with the latter type of regulation revealed an increase in the IL-1 beta TNF alpha combined with a higher heat tolerance. The subjects with the former type of regulation revealed a lower heat tolerance. The increase in the alpha2-macroglobulin appeared to be a most typical acute phase response of the human body to hyperthermia.
Acute-Phase Reaction , Autonomic Nervous System/physiology , Hot Temperature , Acute-Phase Proteins/biosynthesis , Adult , Blood Pressure , Body Temperature , Exercise Test , Heart Rate , Humans , Interleukin-1/biosynthesis , Male , Parasympathetic Nervous System/physiology , Sympathetic Nervous System/physiology , Tumor Necrosis Factor-alpha/biosynthesis
Single-shot hyperthermia of healthy young men in climate chamber up to rectal temperature of 39.5 degrees C over 75-110 minutes revealed variable heat resistance. Individuals with prevalent sympathetic vegetative regulation were more resistant to overheating, if compared to those with prevalent parasympathetic one. Endocrine group peculiarities were observed, the examinees with prevalent parasympathetic vegetative regulation demonstrated more drastic changes of some biochemical parameters.
Fever/blood , Health Status , Occupational Diseases/diagnosis , Adult , Fever/immunology , Humans , Hydrocortisone/blood , Male , Time Factors
