Long noncoding RNA uc.230/CUG-binding protein 1 axis sustains intestinal epithelial homeostasis and response to tissue injury

Circular RNA Cdr1as inhibits proliferation and delays injury-induced regeneration of the intestinal epithelium

“…To determine the involvement of secreted factors originating from stromal or immune cells of the gut in regulation of the intestinal mucosal growth after Cdr1as deletion, we examined the growth of intestinal organoids generated from Cdr1as -/mice. Using methods described previously (8,24), intestinal organoids were derived from proliferating crypts of the mucosa of the mouse small intestine. By 3 days after initial culture, the structures of intestinal organoids from both littermates and Cdr1as -/mice consisted of multiple buds and cells (Figure 4A).…”

“…The recombined Lenti-Cdr1as vector was custom made by AMSBIO, in which intron-Cdr1as locusintron/GFP expression was under the control of the suCMV promoter (24). Lenti-Cdr1as and Lenti-Con vectors were packaged in lentiviral production cells, concentrated by ultracentrifugation, resuspended in PBS, and used to increase Cdr1as in vitro and ex vivo, as described previously (8,24).…”

“…The repair of the intestinal mucosa after injury is a complex process consisting of epithelial restitution characterized by stages of cell spreading and migration into the wounded area, replacement of lost cells by cell proliferation, and epithelial remodeling via differentiation (3)(4)(5). Successful repair of wounds and ulcers requires that intestinal epithelial cells (IECs) change gene expression patterns to modulate cell survival, migration, proliferation, and differentiation (6)(7)(8). Gut mucosal injury occurs commonly in patients with inflammatory bowel disease (IBD) and critical surgical disorders such as sepsis, trauma, and massive thermal injuries (9)(10)(11).…”

Control of Paneth cell function by HuR regulates gut mucosal growth by altering stem cell activity

“…The culture medium and fetal bovine serum were purchased from Invitrogen and biochemical reagents were from Sigma-Aldrich. Isolation and culture of primary enterocytes were conducted following the method described previously ( Yu et al, 2020b ; Yu et al, 2022 ). Briefly, primary crypts were released from the small intestinal mucosa of mice; isolated crypts were mixed with Matrigel (Corning) and cultured in mouse IntestiCult organoid growth medium (Stemcell technology).…”

“…Total RNA was isolated by using the RNeasy mini kit (QIAGEN) and used in RT and quantitative (Q)-PCR amplification reactions as described ( Zhuang et al, 2013 ; Yu et al, 2022 ). Q-PCR analysis was performed using Step-one-plus Systems with specific primers, probes, and software (Applied Biosystems).…”