Journal
NUCLEIC ACIDS RESEARCH
Volume 29, Issue 19, Pages 3975-3981Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/29.19.3975
Keywords
-
Categories
Funding
- NIGMS NIH HHS [R01 GM047110, GM47110] Funding Source: Medline
Ask authors/readers for more resources
To study double-strand break (DSB)-induced mutations in mammalian chromosomes, we stably transfected thymidine kinase (tk)-deficient mouse fibroblasts with a DNA substrate containing a recognition site for yeast endonuclease I-Scel embedded within a functional tk gene. Cells were then electroporated with a plasmid expressing endonuclease I-Scel to induce a DSB, and clones that had lost tk function were selected. In a previous study of DSB-induced tk-deficient clones, we found that similar to8% of recovered tk mutations involved the capture of one or more DNA fragments at the DSB site. Almost half of the DNA capture events involved the I-Scel expression plasmid, and several events involved retrotransposable elements. To learn whether only certain DNA sequences or motifs are efficiently captured, in the current work we electroporated an I-Scel expression plasmid along with Haelll fragments of phi X174 genomic DNA. We report that 18 out of 132 tk-deficient clones recovered had captured DNA fragments, and 14 DNA capture events involved one or more fragments of phi X174 DNA. Microhomology existed at most junctions between phi X174 DNA and genomic sequences. Our work suggests that virtually any extrachromosomal DNA molecule may be recruited for the patching of DSBs in a mammalian genome.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available