The YTHDF1 Knockout IPEC-J2 Cell Line is a CRISPR/Cas9-edited intestinal epithelial cell model featuring targeted disruption of the YTHDF1 gene in the porcine IPEC-J2 cell line. This non-transformed, non-tumorigenic cell line provides a physiologically relevant system for studying m6A?dependent translational regulation in intestinal biology. YTHDF1 gene knockout is achieved via CRISPR/Cas9?mediated genome editing, yielding a loss?of?function model suitable for investigating the functional roles of this m6A reader protein.
The IPEC-J2 cell line was originally derived from the jejunum of a neonatal unsuckled piglet and retains key characteristics of primary intestinal epithelial cells, including microvilli formation, tight junction assembly, and vectorial transport functions. As a non?transformed cell line, IPEC-J2 maintains barrier integrity and polarity, making it an established in vitro platform for studies of epithelial permeability, host?pathogen interactions, and nutrient absorption. Its porcine origin further enables translational research relevant to both human gastrointestinal disorders and swine infectious diseases.
YTHDF1 functions as a cytoplasmic reader of N6?methyladenosine (m6A) modifications on mRNA. It selectively binds m6A?modified transcripts and accelerates their cap?dependent translation by recruiting translation initiation factors, notably components of the eIF3 complex such as EIF3A. This enhanced translation primarily affects transcripts encoding proliferative and pro?survival proteins, including MYC, CCND1, and VEGFA. YTHDF1 expression is positively regulated by transcription factors MYC and HIF1A, and its activity depends on m6A deposition by the METTL3/METTL14 methyltransferase complex and is counterbalanced by erasers FTO and ALKBH5. Through its downstream effectors, YTHDF1 influences tight junction constituents (TJP1, CLDN1, OCLN) and inflammatory mediators (IL6, TNF), thereby connecting epitranscriptomic control to epithelial barrier function and innate immune signaling.
In IPEC-J2 cells, disruption of YTHDF1 is anticipated to reduce translation of m6A?modified mRNAs, leading to impaired cell proliferation and compromised epithelial barrier integrity. This knockout model enables dissection of m6A?dependent post?transcriptional regulation in the intestinal epithelium, especially under pathophysiological conditions such as heat stress?induced injury or viral challenge. For example, the model can be used to investigate how YTHDF1 influences the cellular response to Porcine Epidemic Diarrhea Virus (PEDV) or how its loss alters the expression of key tight junction proteins and inflammatory cytokines, providing mechanistic insights into intestinal barrier dysfunction and inflammatory bowel disease.
Researchers can employ this YTHDF1 knockout cell line for a broad range of translational and basic studies. Typical applications include profiling transcriptome?wide m6A distribution via m6A?seq, validating target transcript translation by polysome profiling, and assessing barrier function through transepithelial electrical resistance (TEER) measurements. Functional assays such as CCK?8 proliferation tests, apoptosis detection, and cytokine ELISAs further enable phenotypic characterization. The model is also suitable for drug target validation in colorectal cancer and for screening compounds that modulate m6A?dependent gene expression in intestinal epithelia. For further information or technical support, please contact Ascent Research.
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