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https://www.readbyqxmd.com/read/29432181/three-classes-of-recurrent-dna-break-clusters-in-brain-progenitors-identified-by-3d-proximity-based-break-joining-assay
#1
Pei-Chi Wei, Cheng-Sheng Lee, Zhou Du, Bjoern Schwer, Yuxiang Zhang, Jennifer Kao, Jeffrey Zurita, Frederick W Alt
We recently discovered 27 recurrent DNA double-strand break (DSB) clusters (RDCs) in mouse neural stem/progenitor cells (NSPCs). Most RDCs occurred across long, late-replicating RDC genes and were found only after mild inhibition of DNA replication. RDC genes share intriguing characteristics, including encoding surface proteins that organize brain architecture and neuronal junctions, and are genetically implicated in neuropsychiatric disorders and/or cancers. RDC identification relies on high-throughput genome-wide translocation sequencing (HTGTS), which maps recurrent DSBs based on their translocation to "bait" DSBs in specific chromosomal locations...
February 20, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27593568/guide-seq-to-detect-genome-wide-double-stranded-breaks-in-plants
#2
Lina Shi, Xiaoqing Tang, Guiliang Tang
Animal and plant cells have repair capabilities to combat DNA damage. DNA damage and repair dynamics can be determined by technologies such as IDLV capture, BLESS, HTGTS, digenome-seq, and GUIDE-seq. Here we highlight GUIDE-seq, a technology used in therapeutics, and envision its application in plants.
October 2016: Trends in Plant Science
https://www.readbyqxmd.com/read/27526713/orientation-specific-rag-activity-in-chromosomal-loop-domains-contributes-to-tcrd-v-d-j-recombination-during-t-cell-development
#3
Lijuan Zhao, Richard L Frock, Zhou Du, Jiazhi Hu, Liang Chen, Michael S Krangel, Frederick W Alt
T cell antigen receptor δ (Tcrd) variable region exons are assembled by RAG-initiated V(D)J recombination events in developing γδ thymocytes. Here, we use linear amplification-mediated high-throughput genome-wide translocation sequencing (LAM-HTGTS) to map hundreds of thousands of RAG-initiated Tcrd D segment (Trdd1 and Trdd2) rearrangements in CD4(-)CD8(-) double-negative thymocyte progenitors differentiated in vitro from bone marrow-derived hematopoietic stem cells. We find that Trdd2 joins directly to Trdv, Trdd1, and Trdj segments, whereas Trdd1 joining is ordered with joining to Trdd2, a prerequisite for further rearrangement...
August 22, 2016: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/27354528/highly-sensitive-and-unbiased-approach-for-elucidating-antibody-repertoires
#4
Sherry G Lin, Zhaoqing Ba, Zhou Du, Yu Zhang, Jiazhi Hu, Frederick W Alt
Developing B lymphocytes undergo V(D)J recombination to assemble germ-line V, D, and J gene segments into exons that encode the antigen-binding variable region of Ig heavy (H) and light (L) chains. IgH and IgL chains associate to form the B-cell receptor (BCR), which, upon antigen binding, activates B cells to secrete BCR as an antibody. Each of the huge number of clonally independent B cells expresses a unique set of IgH and IgL variable regions. The ability of V(D)J recombination to generate vast primary B-cell repertoires results from a combinatorial assortment of large numbers of different V, D, and J segments, coupled with diversification of the junctions between them to generate the complementary determining region 3 (CDR3) for antigen contact...
July 12, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27031497/detecting-dna-double-stranded-breaks-in-mammalian-genomes-by-linear-amplification-mediated-high-throughput-genome-wide-translocation-sequencing
#5
Jiazhi Hu, Robin M Meyers, Junchao Dong, Rohit A Panchakshari, Frederick W Alt, Richard L Frock
Unbiased, high-throughput assays for detecting and quantifying DNA double-stranded breaks (DSBs) across the genome in mammalian cells will facilitate basic studies of the mechanisms that generate and repair endogenous DSBs. They will also enable more applied studies, such as those to evaluate the on- and off-target activities of engineered nucleases. Here we describe a linear amplification-mediated high-throughput genome-wide sequencing (LAM-HTGTS) method for the detection of genome-wide 'prey' DSBs via their translocation in cultured mammalian cells to a fixed 'bait' DSB...
May 2016: Nature Protocols
https://www.readbyqxmd.com/read/26873106/transcription-associated-processes-cause-dna-double-strand-breaks-and-translocations-in-neural-stem-progenitor-cells
#6
Bjoern Schwer, Pei-Chi Wei, Amelia N Chang, Jennifer Kao, Zhou Du, Robin M Meyers, Frederick W Alt
High-throughput, genome-wide translocation sequencing (HTGTS) studies of activated B cells have revealed that DNA double-strand breaks (DSBs) capable of translocating to defined bait DSBs are enriched around the transcription start sites (TSSs) of active genes. We used the HTGTS approach to investigate whether a similar phenomenon occurs in primary neural stem/progenitor cells (NSPCs). We report that breakpoint junctions indeed are enriched around TSSs that were determined to be active by global run-on sequencing analyses of NSPCs...
February 23, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/25503383/genome-wide-detection-of-dna-double-stranded-breaks-induced-by-engineered-nucleases
#7
Richard L Frock, Jiazhi Hu, Robin M Meyers, Yu-Jui Ho, Erina Kii, Frederick W Alt
Although great progress has been made in the characterization of the off-target effects of engineered nucleases, sensitive and unbiased genome-wide methods for the detection of off-target cleavage events and potential collateral damage are still lacking. Here we describe a linear amplification-mediated modification of a previously published high-throughput, genome-wide, translocation sequencing (HTGTS) method that robustly detects DNA double-stranded breaks (DSBs) generated by engineered nucleases across the human genome based on their translocation to other endogenous or ectopic DSBs...
February 2015: Nature Biotechnology
https://www.readbyqxmd.com/read/23611285/translocations-in-normal-b-cells-and-cancers-insights-from-new-technical-approaches
#8
REVIEW
Roberto Chiarle
Chromosomal translocations are recurrent genetic events that define many types of cancers. Since their first description several decades ago as defining elements in cancer cells, our understanding of the mechanisms that determine their formation as well as their implications for cancer progression and therapy has remarkably progressed. Chromosomal translocations originate from double-strand breaks (DSBs) that are brought into proximity in the nuclear space and joined inappropriately by DNA-repair pathways. The frequency and pattern of translocations are influenced by perturbations of any of these events...
2013: Advances in Immunology
https://www.readbyqxmd.com/read/21962511/genome-wide-translocation-sequencing-reveals-mechanisms-of-chromosome-breaks-and-rearrangements-in-b-cells
#9
Roberto Chiarle, Yu Zhang, Richard L Frock, Susanna M Lewis, Benoit Molinie, Yu-Jui Ho, Darienne R Myers, Vivian W Choi, Mara Compagno, Daniel J Malkin, Donna Neuberg, Stefano Monti, Cosmas C Giallourakis, Monica Gostissa, Frederick W Alt
Whereas chromosomal translocations are common pathogenetic events in cancer, mechanisms that promote them are poorly understood. To elucidate translocation mechanisms in mammalian cells, we developed high-throughput, genome-wide translocation sequencing (HTGTS). We employed HTGTS to identify tens of thousands of independent translocation junctions involving fixed I-SceI meganuclease-generated DNA double-strand breaks (DSBs) within the c-myc oncogene or IgH locus of B lymphocytes induced for activation-induced cytidine deaminase (AID)-dependent IgH class switching...
September 30, 2011: Cell
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