Chapters authored
CRISPR Technology: Emerging Tools of Genome Editing and Protein Detection
CRISPR technology has seen rapid development in applications ranging from genomic and epigenetic changes to protein identification throughout the last decade. The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) protein systems have transformed the ability to edit, control the genomic nucleic acid and non-nucleic acid target such as detection of proteins. CRISPR/Cas systems are RNA-guided endonucleases exhibiting distinct cleavage activities deployed in the development of analytical techniques. Apart from genome editing technology, CRISPR/Cas has also been incorporated in amplified detection of proteins, transcriptional modulation, cancer biomarkers, and rapid detection of POC (point of care) diagnostics for various diseases such as Covid-19. Current protein detection methods incorporate sophisticated instrumentation and extensive sensing procedures with less reliable, quantitative, and sensitive detection of proteins. The precision and sensitivity brought in by CRISPR-dependent detection of proteins will ensure the elimination of current impediments. CRISPR-based amplification strategies have been used for accurate estimation of proteins including aptamer-based assay, femtomolar detection of proteins in living cells, immunoassays, and isothermal proximal assay for high throughput. The chapter will provide a comprehensive summary of key developments in emerging tools of genome editing and protein detection deploying CRISPR technology, and its future perspectives will be discussed.
Part of the book: Molecular Cloning
Epigenetics: Science of Changes without Change in DNA Sequences
The mechanisms for epigenetic modifications include modification of histone proteins or modifications of the DNA itself (not affecting the DNA sequence). These include acetylation, methylation, phosphorylation, SUMOylation, ubiquitylation, etc. For example, DNA methylation (cytosine methylation) or histone acetylation (lysine acetylation). Recent studies have indicated that the activity of non-coding RNAs, such as microRNAs, long non-coding RNAs, and small interfering RNAs also affects epigenetic mechanisms. In a genome, the collection of all the modifications that regulate gene expression is called its epigenome. Improper occurrence of the epigenetic mechanisms can lead to deleterious health and behavioral effects. For instance, the most studied epigenetic modification is DNA hypermethylation, which leads to the silencing of antitumorigenic genes, and this has been shown to cause cancer. Various techniques are employed for DNA methylation profiling such as pyrosequencing, bisulfite-PCR, ChIP seq (Chromatin Immunoprecipitation), bisulfite seq, and specialized RNA seq. This chapter will introduce epigenetics, describe the different epigenetic mechanisms, and discuss in brief how to study these mechanisms and their effects on the plant as well as human health.
Part of the book: Modifications in Biomacromolecules