scholarly journals Concatenation of Transgenic DNA: Random or Orchestrated?

Genes ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1969
Author(s):  
Alexander Smirnov ◽  
Nariman Battulin
Keyword(s):  
Dna Repair ◽  
Gene Silencing ◽  
Tandem Repeats ◽  
Routine Procedure ◽  
Dna Integration ◽  
Pronuclear Microinjection ◽  
Transgenic Organisms ◽  
Dna Injection ◽  
Repair Pathways

Generation of transgenic organisms by pronuclear microinjection has become a routine procedure. However, while the process of DNA integration in the genome is well understood, we still do not know much about the recombination between transgene molecules that happens in the first moments after DNA injection. Most of the time, injected molecules are joined together in head-to-tail tandem repeats—the so-called concatemers. In this review, we focused on the possible concatenation mechanisms and how they could be studied with genetic reporters tracking individual copies in concatemers. We also discuss various features of concatemers, including palindromic junctions and repeat-induced gene silencing (RIGS). Finally, we speculate how cooperation of DNA repair pathways creates a multicopy concatenated insert.

scholarly journals Disintegration promotes proto-spacer integration by the Cas1-Cas2 complex

2020 ◽  
Author(s):  
Chien-Hui Ma ◽  
Kamyab Javanmardi ◽  
Ilya J. Finkelstein ◽  
Makkuni Jayaram
Keyword(s):  
Dna Repair ◽  
Streptococcus Pyogenes ◽  
Target Site ◽  
Dna Transposition ◽  
Insertion Event ◽  
Dna Integration ◽  
Biochemical Investigation ◽  
Target Sites ◽  
Repair Pathways ◽  
Integration Event

Abstract‘Disintegration’—the reversal of transposon DNA integration at a target site—is regarded as an abortive off-pathway reaction. Here we challenge this view with a biochemical investigation of the mechanism of protospacer insertion by the Streptococcus pyogenes Cas1-Cas2 complex, which is mechanistically analogous to DNA transposition. In supercoiled target sites, the predominant outcome is the disintegration of one-ended insertions that fail to complete the second integration event. In linear target sites, one-ended insertions far outnumber complete proto-spacer insertions. The second insertion event is most often accompanied by disintegration of the first, mediated either by the 3’-hydroxyl exposed during integration or by water. One-ended integration intermediates may mature into complete spacer insertions via DNA repair pathways that are also involved in transposon mobility. We propose that disintegration-promoted integration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity, and perhaps in other analogous transposition reactions.

scholarly journals Disintegration promotes protospacer integration by the Cas1-Cas2 complex

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Chien-Hui Ma ◽  
Kamyab Javanmardi ◽  
Ilya J Finkelstein ◽  
Makkuni Jayaram
Keyword(s):  
Dna Repair ◽  
Streptococcus Pyogenes ◽  
Target Site ◽  
Dna Transposition ◽  
Insertion Event ◽  
Dna Integration ◽  
Biochemical Investigation ◽  
Target Sites ◽  
Repair Pathways ◽  
Integration Event

‘Disintegration’—the reversal of transposon DNA integration at a target site—is regarded as an abortive off-pathway reaction. Here, we challenge this view with a biochemical investigation of the mechanism of protospacer insertion, which is mechanistically analogous to DNA transposition, by the Streptococcus pyogenes Cas1-Cas2 complex. In supercoiled target sites, the predominant outcome is the disintegration of one-ended insertions that fail to complete the second integration event. In linear target sites, one-ended insertions far outnumber complete protospacer insertions. The second insertion event is most often accompanied by the disintegration of the first, mediated either by the 3′-hydroxyl exposed during integration or by water. One-ended integration intermediates may mature into complete spacer insertions via DNA repair pathways that are also involved in transposon mobility. We propose that disintegration-promoted integration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity, and perhaps in other analogous transposition reactions.

scholarly journals Targeting DNA Repair and Chromatin Crosstalk in Cancer Therapy

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 381
Author(s):  
Danielle P. Johnson ◽  
Mahesh B. Chandrasekharan ◽  
Marie Dutreix ◽  
Srividya Bhaskara
Keyword(s):  
Dna Repair ◽  
Cancer Therapy ◽  
Therapeutic Intervention ◽  
Synthetic Lethality ◽  
Checkpoint Proteins ◽  
Cellular Processes ◽  
Repair Pathways ◽  
Made In

Aberrant DNA repair pathways that underlie developmental diseases and cancers are potential targets for therapeutic intervention. Targeting DNA repair signal effectors, modulators and checkpoint proteins, and utilizing the synthetic lethality phenomena has led to seminal discoveries. Efforts to efficiently translate the basic findings to the clinic are currently underway. Chromatin modulation is an integral part of DNA repair cascades and an emerging field of investigation. Here, we discuss some of the key advancements made in DNA repair-based therapeutics and what is known regarding crosstalk between chromatin and repair pathways during various cellular processes, with an emphasis on cancer.

17α-ethynilestradiol and tributyltin mixtures modulates the expression of NER and p53 DNA repair pathways in male zebrafish gonads and disrupt offspring embryonic development

2018 ◽  
Vol 95 ◽  
pp. 1008-1018 ◽  
Author(s):  
Joana Soares ◽  
Teresa Neuparth ◽  
Angeliki Lyssimachou ◽  
Daniela Lima ◽  
Ana André ◽  
...  
Keyword(s):  
Dna Repair ◽  
Embryonic Development ◽  
Repair Pathways

Investigation on modulation of DNA repair pathways in Chinese MJD patients

2018 ◽  
Vol 71 ◽  
pp. 267.e5-267.e6 ◽  
Author(s):  
Chunrong Wang ◽  
Zhao Chen ◽  
Huirong Peng ◽  
Yun Peng ◽  
Xin Zhou ◽  
...  
Keyword(s):  
Dna Repair ◽  
Repair Pathways

scholarly journals DNA repair pathways as targets for cancer therapy

2008 ◽  
Vol 8 (3) ◽  
pp. 193-204 ◽  
Author(s):  
Thomas Helleday ◽  
Eva Petermann ◽  
Cecilia Lundin ◽  
Ben Hodgson ◽  
Ricky A. Sharma
Keyword(s):  
Dna Repair ◽  
Cancer Therapy ◽  
Repair Pathways

Modulating DNA Repair Pathways to Improve Precision Genome Engineering

2017 ◽  
Vol 13 (2) ◽  
pp. 389-396 ◽  
Author(s):  
Katherine S. Pawelczak ◽  
Navnath S. Gavande ◽  
Pamela S. VanderVere-Carozza ◽  
John J. Turchi
Keyword(s):  
Dna Repair ◽  
Genome Engineering ◽  
Repair Pathways
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