Humans ; Multiple Myeloma/genetics* ; Karyotyping ; Translocation, Genetic/genetics*

Large-scale genetic manipulation of the genome refers to the genetic modification of large fragments of DNA using knockout, integration and translocation. Compared to small-scale gene editing, large-scale genetic manipulation of the genome allows for the simultaneous modification of more genetic information, which is important for understanding the complex mechanisms such as multigene interactions. At the same time, large-scale genetic manipulation of the genome allows for larger-scale design and reconstruction of the genome, and even the creation of entirely new genomes, with great potential in reconstructing complex functions. Yeast is an important eukaryotic model organism that is widely used because of its safety and easiness of manipulation. This paper systematically summarizes the toolkit for large-scale genetic manipulation of the yeast genome, including recombinase-mediated large-scale manipulation, nuclease-mediated large-scale manipulation, de novo synthesis of large DNA fragments and other large-scale manipulation tools, and introduces their basic working principles and typical application cases. Finally, the challenges and developments in large-scale genetic manipulation are presented.
DNA ; Gene Editing ; Genetic Engineering ; Saccharomyces cerevisiae/genetics* ; Translocation, Genetic

Chromosomes, Human ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Translocation, Genetic

Apparently balanced chromosomal structural rearrangements are known to cause male infertility and account for approximately 1% of azoospermia or severe oligospermia. However, the underlying mechanisms of pathogenesis and etiologies are still largely unknown. Herein, we investigated apparently balanced interchromosomal structural rearrangements in six cases with azoospermia/severe oligospermia to comprehensively identify and delineate cryptic structural rearrangements and the related copy number variants. In addition, high read-depth genome sequencing (GS) (30-fold) was performed to investigate point mutations causative of male infertility. Mate-pair GS (4-fold) revealed additional structural rearrangements and/or copy number changes in 5 of 6 cases and detected a total of 48 rearrangements. Overall, the breakpoints caused truncations of 30 RefSeq genes, five of which were associated with spermatogenesis. Furthermore, the breakpoints disrupted 43 topological-associated domains. Direct disruptions or potential dysregulations of genes, which play potential roles in male germ cell development, apoptosis, and spermatogenesis, were found in all cases (n = 6). In addition, high read-depth GS detected dual molecular findings in case MI6, involving a complex rearrangement and two point mutations in the gene DNAH1. Overall, our study provided the molecular characteristics of apparently balanced interchromosomal structural rearrangements in patients with male infertility. We demonstrated the complexity of chromosomal structural rearrangements, potential gene disruptions/dysregulation and single-gene mutations could be the contributing mechanisms underlie male infertility.
Azoospermia/genetics* ; Chromosome Aberrations ; Humans ; Infertility, Male/genetics* ; Male ; Oligospermia/genetics* ; Translocation, Genetic

Humans ; Leukemia, Myeloid, Acute/genetics* ; Myeloid-Lymphoid Leukemia Protein/genetics* ; Oncogene Proteins, Fusion/genetics* ; Translocation, Genetic

Adult ; Gene Deletion ; Humans ; Infertility, Male/genetics* ; Male ; Membrane Proteins/genetics* ; Teratozoospermia/genetics* ; Translocation, Genetic

Karyotypes were prepared from peripheral blood leukocytes in 18 couples with his-tories of habitual abortions. The Standard chromosome analysis and G-banding techniques were studied. The abnormal karyotypes seen were one case with 20% of 45, XX, -14, -15, t(14/15), one case of 46, XY/45, XY, -21 mosaicism, one case of 45, XX, -14, -21, t(14/21), one case of 46, XX/45, XO mosaicism and one case of 46, XYq+. Many other types of chromosomal abnormalities from many reports in couples with spontaneous abortions are discussed.
Abortion, Habitual/genetics* ; Adult ; Chromosome Aberrations* ; Female ; Human ; Leukocytes/ultrastructure ; Male ; Mosaicism ; Pregnancy ; Translocation (Genetics)

Chromosomal translocation is a kind of common chromosomal abnormality. The carriers with chromosomal translocation could have more chance of normal pregnancy with the help of fluorescence in situ hybridization(FISH). This is a review aimed at analyzing the meiosis types of the translocation chromosome. The strategy of preimplantation genetic diagnosis for the carriers with chromosomal translocation is also discussed.
Humans ; In Situ Hybridization, Fluorescence ; methods ; Meiosis ; genetics ; Preimplantation Diagnosis ; methods ; Translocation, Genetic ; genetics

OBJECTIVETo analyze the meiotic segregation results of male reciprocal chromosome translocation by fluorescence in situ hybridization (FISH).

METHODSMulti-color FISH using 3 combined probes located in any 3 chromosome segments on both sides of two breakpoints was performed on the de-condensed sperm head to analyze the sperm chromosomal contents and segregation patterns.

RESULTSFour male reciprocal translocation carriers were included in the study, with the karyotypes of 46, XY, t(2;18) (p16; q23); 46, XY, t(4;6) (q34;q21); 46, XY, t(8;13) (q23;q21) and 46, XY, t(4;5) (4q31;5q13), respectively. The results showed that 4 carriers had different proportions of various segregated spermatozoa. The spermatozoa of alternate, adjacent-1, adjacent-2, 3:1, non-disjunction in meiosis II, and 4:0 or diploidy accounted for 27.1%-49.4%, 26.9%-37.6%, 2.7%-15.7%, 8.6%-32.7%, 0.2%-1.9%, and 0.1%-0.4%, respectively.

CONCLUSIONFor each-reciprocal translocation carrier seems to have a particular meiotic segregation results, FISH analysis on sperm head should be done for each carrier in order to provide an accurate genetic counseling.

Chromosome Breakage ; Heterozygote ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Meiosis ; genetics ; Spermatozoa ; metabolism ; Translocation, Genetic ; genetics

Adult ; Azoospermia ; genetics ; Chromosomes, Human, Pair 13 ; genetics ; Chromosomes, Human, Pair 14 ; genetics ; Humans ; Hypogonadism ; genetics ; Male ; Translocation, Genetic