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. 2021 Dec 2;101(5):551–558. doi: 10.1177/00220345211049984

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© International Association for Dental Research and American Association for Dental, Oral, and Craniofacial Research 2021

This article is distributed under the terms of the Creative Commons Attribution 4.0 License (https://creativecommons.org/licenses/by/4.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 4. — rs11084095 and rs4284742 have allele-specific effects on luciferase activity. The 65-bp DNA sequence up- and downstream of rs11084095 and the 75-bp DNA sequence spanning rs4284742 showed allele-specific enhancer activity in HeLa cells. (A) In the background of the common rs11084095 G-allele, luciferase activity was 9.9-fold increased (P = 0.015) versus the rare A-allele, which showed no upregulation. The difference between alleles was significant with P = 0.013. (B) The common rs4284742 G-allele increased luciferase activity 13-fold (P = 0.008) as compared with the rare A-allele, which increased the activity 4-fold (P = 0.002). The differences between alleles was significant with P = 0.010. (C) Both alleles of rs34984145 showed significant enhancer activity in HeLa cells. The rare T-allele showed stronger upregulation of 4.2-fold (P = 0.0001) versus the common A-allele, which showed an upregulation of 2.3-fold (P = 0.002). The difference between alleles was not significant. (D) The haplotype block at rs4284742 has a cis-regulatory effect on SIGLEC5 expression. CRISPRa of the genomic region at rs4284742 induced SIGLEC5 expression as compared with the scrambled gRNA. gRNA 1 (chr19:52131631-52131649), gRNA 2 (chr19:52131441-52131459), and gRNA 3 (chr19:52131754-52131773) induced SIGLEC5 expression 380-fold (P = 0.00008), 137-fold (P = 0.03), and 80-fold (P = 0.0009), respectively (positions are given for genome build GRCh37/hg19). (E) CRISPRa of the genomic region at rs11084095 did not significantly induce SIGLEC5 expression (gRNA 1: chr19:52126982-52127000, gRNA 2: chr19:52127105-52127123, gRNA 3: 2126912-52126930). Error bars indicate 95% CI. *P < .05. **P < .01. ***P < .001.