Preface.- Part 1. Improved DNA polymerases and new DNA and DNA/RNA ligases useful in RCA.- 1. Improvement of φ29 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.
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).
This book covers the latest developments in rolling circle amplification (RCA) technology with applications in clinical diagnostic tests and molecular medicine. Topics covered include new enzymes useful in RCA, techniques involving RCA for enhanced signal amplification, novel RCA diagnostics, sensors for expediting RCA detection, and prospective RCA-based therapeutics. This is a valuable book for university professors and students in the field of biomedical engineering and biomolecular pharmacology as well as R&D managers of biotechnology and biopharmaceutical companies.
Specifically, this book:
Reviews prospective RCA-based therapeutics, including RCA-derived DNA nanoparticles that strongly bind to cancer cells
Expands readers’ understanding of sensor systems for expediting detection of RCA products by using probe-tagged magnetic nanobeads
Maximizes reader insights into novel RCA diagnostics, such as PNA openers-assisted RCA for detection of single target cells and in situ RCA diagnosis of cancer cells and malignant tissues
Presents innovative methods for quasi-exponential enhancement of RCA-generated signals, such as nicking enzyme-assisted cascade RCA and RCA coupled with loop-mediated amplification
Advance Praise for Rolling Circle Amplification (RCA):
“This book provides a badly needed compendium of innovative RCA methods and applications. It should help further increase the community of scientists that have employed RCA in research and diagnostic programs.”
— Charles Cantor, Professor Emeritus of Biomedical Engineering, Boston University Executive Director, Retrotope Inc. (USA)
“In this new book Vadim Demidov has assembled an enticing menu of articles that illustrate the evolution of the RCA field, including improved protein parts for building superior DNA nanomachines, enhanced modalities of amplification and detection, diagnostic applications, and even a sampling of potential therapeutic applications. The reader will appreciate that while RCA has come of age, there is no lack of exciting surprises, turns, and twists in the continuing evolution of the technology.”
— Paul Lizardi, Professor of Pathology, Yale University School of Medicine (retired) Investigator, University of Granada, Spain, President, PetaOmics, Inc., San Marcos, Texas.