Paul F Kisak

CRISPR Technology Paul F Kisak

CRISPR is an acronym for Clustered Regularly Interspaced Short Palindromic Repeats which are segments of prokaryotic DNA containing short, repetitive base sequences. These play a key role in a bacterial defense system and form the basis of a genome editing technology known as CRISPR/Cas9 that allows permanent modification of genes within organisms. In a palindromic repeat, the sequence of nucleotides is the same in both directions. Each repetition is followed by short segments of spacer DNA from previous exposures to foreign DNA (e.g., a virus or plasmid). Small clusters of cas (CRISPR-associated system) genes are located next to CRISPR sequences. The CRISPR/Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages that provides a form of acquired immunity. RNA harboring the spacer sequence helps Cas proteins recognize and cut exogenous DNA. Other RNA-guided Cas proteins cut foreign RNA. CRISPRs are found in approximately 40% of sequenced bacterial genomes and 90% of sequenced archaea. A simple version of the CRISPR/Cas system, CRISPR/Cas9, has been modified to edit genomes. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added. The Cas9-gRNA complex corresponds with the CAS III crRNA complex in the above diagram. CRISPR/Cas genome editing techniques have many potential applications, including medicine and crop seed enhancement. The use of CRISPR/Cas9-gRNA complex for genome editing was the AAAS's choice for breakthrough of the year in 2015. Bioethical concerns have been raised about the prospect of using CRISPR for germline editing. This book is designed to be a state of the art, superb academic reference work and provide an overview of the topic and give the reader a structured knowledge to familiarize yourself with the topic at the most affordable price possible. The accuracy and knowledge is of an international viewpoint as the edited articles represent the inputs of many knowledgeable individuals and some of the most current knowledge on the topic, based on the date of publication.