Abstract
Mutations and single base pair polymorphisms in various genes have been associated with increased susceptibility to inflammatory bowel disease (IBD). We have created a series of rat strains carrying targeted genetic alterations within three IBD susceptibility genes: Nod2, Atg16l1, and Il23r, using CRISPR/Cas9 genome editing technology. Knock-out alleles and alleles with known human susceptibility polymorphisms were generated on three different genetic backgrounds: Fischer, Lewis and Sprague Dawley. The availability of these rat models will contribute to our understanding of the basic biological roles of these three genes as well as provide new potential IBD animal models.
Highlights
Inflammatory bowel disease (IBD) refers to a group of diseases involving chronic inflammation of the gastrointestinal tract (GI)
Resulting pups were initially screened by PCR genotyping using three sets of primers designed to the region targeted by each gRNA
DNA from animals with evidence of a mutation based on this initial PCR screen, was further analyzed by nucleotide sequence analysis to characterize the exact nature of the mutation
Summary
Inflammatory bowel disease (IBD) refers to a group of diseases involving chronic inflammation of the gastrointestinal tract (GI). The main forms of IBD are Crohn’s disease (CD) and ulcerative colitis (UC). CD can affect any part of the GI, and affect extra-intestinal organ and tissue systems (Xavier and Podolsky 2007). UC, by comparison, is restricted to the colon and rectum, commonly manifests and spreads from a focal region, and involves only the mucosal layer of the GI. Both diseases have the potential to wreak havoc upon the GI system, cause life-threatening perforation and stricture of the gut, and result in life-long physical and emotional turmoil.
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