Rolling Circle Amplification (RCA)

Vadim V. Demidov, Ph.D., is a Senior Analyst in the Biotechnology & Pharmaceuticals Group at Global Prior Art, Inc. (Boston, USA), an intellectual property research and analysis firm. Dr. Demidov received his M.S. degree in Physical and Chemical Engineering from Moscow Institute of Physics and Technology (MIPT, a leading Russian technical university, also known as “the Russian MIT”) and his Ph.D. degree in Biophysics from the Institute of Molecular Genetics of the Russian Academy of Sciences (IMGRAS) and MIPT.Before joining the Global Prior Art company in 2008, he worked for almost 30 years in academia and industry worldwide. Dr. Demidov is well known in the molecular biology and biotechnology field for his innovative studies related to peptide nucleic acid (PNA). He has introduced the use of PNA openers for detection of specific sequences within DNA duplexes under non-denaturing conditions, with some of these applications involving rolling circle amplification (RCA).

Preface.- Part 1. Improved DNA polymerases and new DNA and DNA/RNA ligases useful in RCA.- 1. Improvement of phi29 DNA polymerase amplification performance by fusion of DNA binding motifs.- 2. Preparation of circular templates by T4 RNA ligase 2 for rolling circle amplification of target microRNAs with high specificity and sensitivity.- 3. Use of DNA CircLigase for direct isothermal detection of microbial mRNAs by RNA-primed rolling circle amplification and preparation of ø29 DNA polymerase not contaminated.- Part 2. RCA-involving techniques with enhanced signal amplification.- 4. Nicking enzyme-assisted branched-chain RCA reaction for cascade DNA amplification.- 5. Combining isothermal amplification techniques: Coupled RCA-LAMP.- Part 3. Emerging RCA diagnostics.- 6. Detection of vascular disease-related single nucleotide polymorphisms in clinical samples using ramified rolling circle amplification.- 7. Ultrasensitive isothermal detection of protein analytes using rolling circle amplification in micro-scale platforms.- 8. Rolling circle amplification with padlock probes for in situ detection of RNA analytes.- 9. PNA-assisted rolling circle amplification for detection of DNA marker sequences in human cells.- 10. Sensor systems with magnetic and optomagnetic readout of rolling circle amplification products.- Part 4. Prospective RCA-based therapeutics.- 11. DNA nanoclews for stimuli-responsive anticancer drug delivery.- 12. RCA-assisted self-assembled DNA origami nano-constructs as vehicles for cellular delivery of diagnostic probes and therapeutic drugs.- 13. DeNAno: a novel multivalent affinity reagent produced by selection of RCA-generated DNA nanoparticle libraries.- Index.