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Human, mouse, and Arabidopsis genes (Duan et al., 2013, Narsai et al., 2007, Sharova et al., 2009, Wang et al., 2007). Insertion of an intron into cDNA constructs containing instability determinants increased their stability in human cells (Zhao and Hamilton, 2007). It was also shown that introns can enhance pre-mRNA 3′ end processing and polyadenylation. This effect was mediated by the splicingIntrons can increase the efficiency of mRNA translationBesides increasing mRNA content by affecting transcription, export, and stability, the presence of introns was also shown to increase the efficiency of mRNA translation in mammals, plants, yeast, and Xenopus (Akua and Shaul, 2013, Bourdon et al., 2001, Curi et al., 2005, Gudikote et al., 2005, Hoshida et al., 2017, Lu and Cullen, 2003, Mascarenhas et al., 1990, Matsumoto et al., 1998, Nott et al., 2003, Nott et al., 2004, Rose, 2004, Samadder et al., 2008). The ability of introns, which areThe role of splicing in the enhancement of gene expression by intronsThere is an ongoing discussion in the IME literature whether splicing itself is essential for the ability of introns to elevate gene expression. This issue is discussed here in light of the above-presented data. Several studies showed that the presence of introns was sufficient to enhance expression even when splicing was prevented (e.g., Damgaard et al., 2008, Kaida et al., 2010, Kwek et al., 2002). These data can be at least partially explained by the splicing-independent functions of U1The significance of intron’s position relative to the sites of transcription and translation initiationExperiments in which the same intron was inserted in different positions of the same gene indicated that the stimulatory impact of introns on transcription depends on their proximity to the promoter (Furger et al., 2002, Rose, 2004). There are several possible reasons for these findings. It is reasonable that Prize winner. The Participant must provide accurate, current and complete information when participating in the Promotion. The Promoter may request a winner, before presenting the Prize, to provide his/her identity and residence information previously provided by the Participant. If a Participant does not truthfully provide all requested Personal Information, the Promoter may determine that they are not eligible to win the Prize. Participant acknowledges that the Promoter can rely on these Terms and Conditions and disqualify a Participant if the Promoter believes the Participant is not acting in conformity with the Terms and Conditions. Promoter may require the Participant to return a Prize or pay its equivalent value if Promoter learns of a Participant’s ineligibility or disqualification after awarding the Prize to such ineligible or disqualified Participant. If for any reason the conduct or operation of this Promotion is interfered with or disrupted in any way (including but not limited to, vandalism, power failures, natural disasters, acts of God, governmental acts or orders or restrictions, local restrictions, civil unrest, strikes, computer bugs or viruses, fraud, tampering, technical failures or any other causes beyond the control of the Promoter), the Promoter reserves the right to cancel, terminate, modify or suspend or recommence the Promotion in whole or in part. Neither party shall be liable to the other party for damages or losses in this case. The Promoter assumes no responsibility for any incorrect or incomplete information which may be communicated in the course of administering this Promotion. The Promoter reserves the right to terminate this Promotion if fraud, technical failures or any other factor beyond Promoter's reasonable control impairs the integrity of the Promotion as determined by the Promoter in its sole discretion. Online entries made with multiple e-mail addresses, under multiple identities or through the use of any other device or artifice to enter multiple times will be deemed invalid. If the winner is an online entrant, the winning online entry will be deemed to have been submitted by the authorized account holder of the e-mail address from which the entry was made. The authorized email account holder is deemed as the natural person who is assigned to an e-mail address by an internet access provider, service provider or other organization that is responsible for assigning e-mail addresses or the domain associated with the submitted e-mail address. The Promoter shall not be responsible for incorrect or inaccurate transcription of entry

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Mimics or inhibitors. H HCT15 cells were transfected with miR-7-5p mimics or control, then treated with Licl or control. The expression of ACVRL1 was detected by western blot and qRT-PCR. I Schematic representation of the KCNQ1OT1 promoter region and the reporter constructs used for the luciferase assay. One reporter contains the KCNQ1OT1 promoter region from −294 to −2022 (Kp2022). The other reporter contains a truncated promoter in which the TCF-1 region was deleted (Kp1080). J The relative luciferase activity was detected in HCT15 cells co-transfected by Kp2022/Kp1080 and β-catenin. K The ChIP assay of β-catenin binding to the TCF site in the KCNQ1OT1 promoter in CRC cell lines. The assay was performed with or without β-catenin antibody. DNA was recovered from immunoprecipitated and nonimmunoprecipitated (input) chromatin and analysed by qRT-PCR. L The expression of miR-7-5p in CRC cells transfected by shKCNQ1OT1. M The predicted binding sites between miR-7-5p and KCNQ1OT1. The relative luciferase activities were detected in HCT15 cells transfected by KCNQ1OT1-WT and KCNQ1OT1-Mut (ns means no significance. *P P P P Full size imageIn order to further verify the molecular mechanism of the inhibition of miR-7-5p by Wnt/β-catenin pathway, we conducted the following explorations. Studies have shown that lncRNA-KCNQ1OT1 transcription is directly regulated by β-catenin in CRC [17]. To investigate whether β-catenin regulates lncRNA-KCNQ1OT1 transcription through association with the KCNQ1OT1 promoter, we selected a TCF-1 binding site that is predicted to be located closest to the transcription start site on KCNQ1OT1. We constructed human KCNQ1OT1 promoter-luciferase reporter plasmids: plasmid Kp2022 contained a 2020-bp fragment from within the KCNQ1OT1 promoter region that included this TCF-1 binding site, and plasmid Kp1080 contained a truncated fragment of the KCNQ1OT1 promoter that did not include this TCF-1 binding site (Fig. 6I). We then examined the effect of co-transfection of a β-catenin expressing vector