1.Tools for large-scale genetic manipulation of yeast genome.
Jieyi LI ; Hanze TONG ; Yi WU
Chinese Journal of Biotechnology 2023;39(6):2465-2484
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
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Gene Editing
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Genetic Engineering
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Saccharomyces cerevisiae/genetics*
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Translocation, Genetic
2.Is t(11;14)(q13;q32) good or bad for newly diagnosed multiple myeloma?
Yang LIU ; Lu GAO ; Yueyun LAI ; Lei WEN ; Wenbing DUAN ; Fengrong WANG ; Ling MA ; Xiaojun HUANG ; Jin LU
Chinese Medical Journal 2023;136(1):96-98
3.Therapy-related acute myeloid leukemia with t(9;22)(q34;q11) and t(16;21)(q24;q22) double translocation: a case report and literature review.
Xiao Yan DONG ; Yu Long LI ; Cheng Ye WU ; Wei CHENG ; Bao Jun SHANG ; Lin ZHANG ; Lin Na CHENG ; Zun Min ZHU
Chinese Journal of Hematology 2019;40(11):956-958
4.Investigation of the genetic etiology in male infertility with apparently balanced chromosomal structural rearrangements by genome sequencing.
Matthew Hoi Kin CHAU ; Ying LI ; Peng DAI ; Mengmeng SHI ; Xiaofan ZHU ; Jacqueline Pui WAH CHUNG ; Yvonne K KWOK ; Kwong Wai CHOY ; Xiangdong KONG ; Zirui DONG
Asian Journal of Andrology 2022;24(3):248-254
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*
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Chromosome Aberrations
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Humans
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Infertility, Male/genetics*
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Male
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Oligospermia/genetics*
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Translocation, Genetic
7.Preimplantation genetic testing for a couple where the husband is affected by osteogenesis imperfecta combined with balanced translocation using karyomapping technique.
Wenbin NIU ; Mingzhu HUO ; Hao SHI ; Yidong LIU
Chinese Journal of Medical Genetics 2021;38(11):1068-1072
OBJECTIVE:
To carry out preimplantation genetic testing (PGT) for a couple where the husband was affected by osteogenesis imperfecta combined with balanced translocation using the karyomapping technique.
METHODS:
Blastocysts were detected using karyomapping, the carrier status of COL1A1 c.760G>A (p.Gly254Arg) variant and the carrier status of the translocated chromosome were analyzed simultaneously.
RESULTS:
For a total of 10 blastocysts, two euploid blastocysts were found to not carry the COL1A1 c.760G>A (p.Gly254Arg) variant but a balanced translocation. After transplanting one of the blastocysts, clinical pregnancy was achieved. Amniocentesis at 18th gestational week and prenatal genetic testing was in keeping with the result of PGT.A healthy female was born at 40+4 weeks gestation.
CONCLUSION
For patients simultaneously carrying genetic variant and balanced chromosomal translocation, PGT can be performed with efficiency by the use of karyomapping method.
Blastocyst
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Female
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Fertilization in Vitro
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Genetic Testing
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Humans
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Osteogenesis Imperfecta/genetics*
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Pregnancy
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Preimplantation Diagnosis
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Spouses
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Translocation, Genetic
8.Advances in preimplantation genetic diagnosis for chromosomal translocation carrier.
Yueqiu TAN ; Guangxiu LU ; Luyun LI
Chinese Journal of Medical Genetics 2002;19(1):76-78
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
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In Situ Hybridization, Fluorescence
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methods
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Meiosis
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genetics
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Preimplantation Diagnosis
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methods
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Translocation, Genetic
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genetics
9.Meiotic segregation results of male reciprocal chromosome translocations.
Xue-feng HUANG ; Shi-quan XIAO ; Qian-jin FEI ; Wei ZHANG ; Li-ya ZHANG ; Xu YANG ; Bi-lv YE
Chinese Journal of Medical Genetics 2007;24(2):217-220
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