Technical Review: Cytogenetic Tools for Studying Mitotic Chromosomes

Abstract

Significant advances in chromosome preparation and other techniques have greatly increased the potential of plant cytogenetics in recent years. Increase in longitudinal resolution using DNA extended fibers as well as new developments in imaging and signal amplification technologies have enhanced the ability of FISH to detect small gene targets. The combination of fluorescence in situ hybridization with immunocytochemistry allows the investigation of cell events, chromosomal rearrangements and chromatin features typical for plant nuclei. Chromosome manipulation techniques using microdissection and flow sorting have accelerated the analysis of complex plant genomes. Together, the different cytogenetic approaches are invaluable for the unravelling of detailed structures of plant chromosomes, which are of utmost importance for the study of genome properties, DNA replication and gene regulation. In this technical review, different cytogenetic approaches are discussed for the analysis of plant chromosomes, with a focus on mitotic chromosomes.

The Formation of Sex Chromosomes in Silene latifolia and S. dioica Was Accompanied by Multiple Chromosomal Rearrangements

“…Recent advances in fluorescence in situ hybridization (FISH) experiments have provided a variety of techniques which can be used to study the structure, dynamics and origin of certain loci, chromosomal arms and/or specific chromosomes (reviewed in Cui et al, 2016;Bačovský et al, 2018;Huber et al, 2018). Previous cytogenetic studies in Silene species were based mainly on physical mapping of satellite rDNA (Široký et al, 2001), repeats and transposable elements (Cermak et al, 2008;Kejnovsky et al, 2009).…”

Molecular genetic testing methodologies in hematopoietic diseases: current and future methods

“…[12][13][14][15][16][17] Some disadvantages are that the karyotype relies on living cells to culture, and yield of viable cells may be limited by chemotherapy exposure or sampling. 18 Additionally, subtle genetic rearrangements may not be detected. This method is generally not applicable to minimal residual disease monitoring in the setting of current technologies.…”

“…This technology has been a powerful driver of diagnostic subtyping, prognosis, and monitoring of diseases in myeloid neoplasms . Some disadvantages are that the karyotype relies on living cells to culture, and yield of viable cells may be limited by chemotherapy exposure or sampling . Additionally, subtle genetic rearrangements may not be detected.…”

Overview of Cytogenetic Technologies

“…It allows the study of fundamental biological questions, such as the nature of inherited syndromes, genomic alterations that are involved in tumorigenesis, and three-dimensional organization of the human genome [2]. Over the years, cytogenetic techniques have been used to unravel the three-dimensional organization of the genome and epigenetic features of higher-order chromatin structure [1,[3][4][5][6][7][8]. Throughout the history, various discoveries and techniques have revolutionized the field of human cytogenetics (Table 1) [1].…”