ABSTRACT
One of the latest methods in modern molecular biology is labeling genomic loci in living cells using fluorescently labeled Cas protein. The NIH Foundation has made the mapping of the 4D nucleome (the three-dimensional nucleome on a timescale) a priority in the studies aimed to improve our understanding of chromatin organization. Fluorescent methods based on CRISPR-Cas are a significant step forward in visualization of genomic loci in living cells. This approach can be used for studying epigenetics, cell cycle, cellular response to external stimuli, rearrangements during malignant cell transformation, such as chromosomal translocations or damage, as well as for genome editing. In this review, we focused on the application of CRISPR-Cas fluorescence technologies as components of multimodal imaging methods for in vivo mapping of chromosomal loci, in particular, attribution of fluorescence signal to morphological and anatomical structures in a living organism. The review discusses the approaches to the highly sensitive, high-precision labeling of CRISPR-Cas components, delivery of genetically engineered constructs into cells and tissues, and promising methods for molecular imaging.
Subject(s)
CRISPR-Cas Systems , Gene Editing , Gene Editing/methods , Genome , Genomics , Microscopy, FluorescenceABSTRACT
Introduction:When a central nervous system disorder (meningitis, encephalitis, hemorrhage, leukemia infltration and other neoplasma) is present, cerebrospinal fluid (CSF) shows various changes that reflected the condition. Therefore it is essential to test CSF. Different types of CSF tests include cell count; cell differentiation; chemistry; immunology; microbiology and molecular biology. CSF cell count and cell differentiation in particular, are crucial in differentiating diagnosing various CNS disorder needing immediate care and in evaluating the treatment. The patient’s prognosis largely depends on how accurate diagnosis was done and how early treatment was provided. There for CSF test require high precision and accuracy. In Mongolia until now 2st and 3st level hospital using manual method for CSF cell count and cell differentiation test. In this test has 2 actual problems, which is depends on the analytical techniques, skills and sample stability specific problem. But in Japan in 2011 newly designed Sysmex XN Series hematology analyser with body fluid mode (CSF,pleural effusion, peritoneal and synovial fluid). On The First Central Hospital of Mongolia In 2013 frst timeinstalled Sysmex XN-2000 hematology analyser andpossible use of body fluid automatic testing methods.Materials and methods:We evaluated the basic assay performance of the body fluid mode on the automated hematology analyzer XN-2000, which is used for analysis of CSF fluid. We compared between the manual method and XN-2000 analysis for nucleated (WBC), mononuclear (MN) and polymorphonuclear (PMN) cells was also randomly studied using 10 CSF samples of inpatient section our hospital.Results:In CSF samples the coeffcient correlation(r) for WBC/µl, MN%, PMN% were respectively 0.83, 0.95 ба 0.95.Discussion:The correlation for MN%, PMN% were between automate and manual method was good, that is similar to the other researchers. Whereas the correlation for WBC/µl slightly low, this was probably correlation relatively weak or show discrepancies. In introduction inscriptive in analysis accuracy can to affect analytical techniques skills, sample stability and specifc many problems. Therefore scientifc studied and proven ability specifcity, sensitivity, reproducibility, quality, personnel low cost and spend less time, automatically Sysmex XN series hematology analyzer is desirable to domesticate an appropriate level of medical laboratories.
