CRISPR/Cas9 technology is one of the most popular methods used for genome editing. It introduces both Cas9 endonuclease and a guide RNA into the cells of interest. The guide RNA is designed to direct the Cas9 endonuclease to a particular site in the genome where it produces a double-strand break (DSB). There are generally two ways to repair the double-stranded break: non-homologous end-joining (NHEJ) or homologous-directed repair (HDR). NHEJ is the main form of repair in mammalian cells. As it is error-prone, repair via the NHEJ pathway allows for insertion, deletion, and loss-of-function mutations which probably result in frameshifts and affect protein expression. In addition to knockout mutations, a template DNA can be introduced to direct HDR and create mutations in the gene of interest. HDR faithfully copies the template sequence to the cut target site.
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In microbiome research (including metagenomics and 16S rRNA sequencing), the terms absolute abundance and relative abundance are frequently encountered. However, what exactly do these terms mean, and why is it important to differentiate between them?
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