A5029353306- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- Bacterial Genetics and Biotechnology
- Genomics and Chromatin Dynamics
- Microtubule and mitosis dynamics
- DNA and Nucleic Acid Chemistry
- Advanced biosensing and bioanalysis techniques
- Genetics, Aging, and Longevity in Model Organisms
- Advanced Fluorescence Microscopy Techniques
- RNA and protein synthesis mechanisms
- RNA Interference and Gene Delivery
University of Wollongong
2018-2023
Illawarra Health and Medical Research Institute
2019-2023
Abstract Limited experimental tools are available to study the consequences of collisions between DNA-bound molecular machines. Here, we repurpose a catalytically inactivated Cas9 (dCas9) construct as generic, novel, targetable protein–DNA roadblock for studying mechanisms underlying enzymatic activities on DNA substrates in vitro . We illustrate broad utility this tool by demonstrating replication fork arrest specifically bound dCas9–guideRNA complex viral, bacterial and eukaryotic forks
Abstract Genome duplication occurs while the template DNA is bound by numerous DNA-binding proteins. Each of these proteins act as potential roadblocks to replication fork and can have deleterious effects on cells. In Escherichia coli, are displaced accessory helicase Rep, a translocase that interacts with replisome. The mechanistic details underlying coordination roadblock removal Rep remain poorly understood. Through real-time fluorescence imaging produced individual E. coli replisomes...
DNA replication occurs on chromosomal while processes such as repair, recombination and transcription continue. However, we have limited experimental tools to study the consequences of collisions between DNA-bound molecular machines. Here, repurpose a catalytically inactivated Cas9 (dCas9) construct fused photo-stable dL5 protein fluoromodule novel, targetable protein-DNA roadblock for studying fork arrest at single-molecule level in vitro well vivo . We find that specifically bound...
ABSTRACT Genome duplication occurs while the template DNA is bound by numerous DNA-binding proteins. Each of these proteins act as potential roadblocks to replication fork and can have deleterious effects on cells. In Escherichia coli , are displaced accessory helicase Rep, a translocase that interacts with replisome. The mechanistic details underlying coordination roadblock removal Rep remain poorly understood. Through real-time fluorescence imaging produced individual E. replisomes...