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Tetsushi Sakuma, Takuya Sakamoto, Takashi Yamamoto
CRISPR-Cas9 enables highly convenient multiplex genome engineering in cultured cells, because it utilizes generic Cas9 nuclease and an easily customizable single-guide RNA (sgRNA) for site-specific DNA double-strand break induction. We previously established a multiplex CRISPR-Cas9 assembly system for constructing an all-in-one vector simultaneously expressing multiple sgRNAs and Cas9 nuclease or other Cas9 variants including FokI-dCas9, which supersedes the wild-type Cas9 with regard to high specificity. In this chapter, we describe a streamlined protocol to design and construct multiplex CRISPR-Cas9 or FokI-dCas9 vectors, to introduce them into cultured cells by lipofection or electroporation, to enrich the genomically edited cells with a transient puromycin selection, to validate the mutation efficiency by Surveyor nuclease assay, and to perform off-target analyses...
2017: Methods in Molecular Biology
Yan Cao, Xinjing Wang, Zhihong Cao, Xiaoxing Cheng
INTRODUCTION: The association between FokI polymorphism of vitamin D receptor (VDR) and tuberculosis (TB) susceptibility has been investigated previously; however, the results were inconsistent and conflicting. In the present study, a meta-analysis was performed to assess the relationship between VDR FokI gene polymorphism and the risk of TB. MATERIAL AND METHODS: Databases including PubMed and Embase were searched for genetic association studies of FokI polymorphism of vitamin D receptor (VDR) and TB...
October 1, 2016: Archives of Medical Science: AMS
Gaye Erten, Muhammed Kalkan, Sema Bilgiç Gazioğlu, Nilgun Akdeniz, Elif Ozkok, Burcak Vural
Objectives. In our study we aimed to determine VDR gene polymorphisms in patients with Behçet's disease (BD) and neuro-Behçet's disease (NBD) in Turkish population. Methods. PBL obtained from 37 patients with BD, 21 patients with NB, and 30 healthy controls were investigated. Genomic DNA was extracted from whole blood using the QIAamp Blood Kit. VDR ApaI (rs7975232), VDR FokI (rs2228570), and VDR TaqI (rs731236) genotyping was performed by real-time polymerase chain reaction with SimpleProbe melting-curve analysis...
2016: Disease Markers
Maimoona Nasreen, Khalid Pervaiz Lone, Saba Khaliq, Shagufta Khaliq
OBJECTIVE: To compare the pattern of Vitamin D receptor (VDR) polymorphisms (Apa I and Fok I) in Type I Diabetes mellitus (T1DM) as cases vs healthy population as control and to investigate the association of VDR polymorphism with vitamin D levels in cases and controls. METHODS: The hypothesis of the study was "VDR gene polymorphisms (Fok 1 and Apa 1) and vitamin D levels are associated with the T1DM". The case-control study was carried out on 44 cases and 44 controls...
October 2016: JPMA. the Journal of the Pakistan Medical Association
Aline Hajj, Rima Chedid, Eliane Chouery, André Megarbané, Marie-Hélène Gannagé-Yared
AIM: To explore the association between VDR polymorphisms and several cardiovascular risk factors and adiponectin. MATERIALS & METHODS: Three-hundred and sixty-nine healthy students were randomly selected. Five VDR polymorphisms were genotyped: BsmI rs1544410; Cdx2 rs11568820; ApaI rs7975232; TaqI rs731236 and FokI rs2228570. BMI, waist circumference (WC), blood pressure, lipid/glycemic profiles and adiponectin were assessed. RESULTS: In men, BsmI, ApaI and TaqI were associated with BMI and WC (p < 0...
October 2016: Pharmacogenomics
Hua Jiang, Zhilin Qin, Shaohui Zong, Maolin He, Xinli Zhan, Zengming Xiao, Qingjun Wei
PURPOSE: To examine the association between Vitamin D receptor (VDR) gene polymorphisms and lumbar disc degeneration (LDD) predisposition. METHODS: A comprehensive literature search was conducted to identify all the relevant studies. The allele/genotype frequencies were extracted from each study. We calculated the pooled odds ratios (ORs) and 95 % confidence intervals (CI) to assess the strength of the association between the VDR gene polymorphisms and LDD risk...
September 9, 2016: European Spine Journal
Dong-Dong Zhao, Dan-Dan Yu, Qiong-Qiong Ren, Bao Dong, Feng Zhao, Ye-Huan Sun
BACKGROUND: As for the association of vitamin D receptor (VDR) gene polymorphisms with susceptibility to pediatric asthma, results of published studies yielded conflicts. A systematic review was conducted on the relationship between childhood asthma and VDR gene polymorphisms, including ApaI (rs7975232), BsmI (rs1544410), FokI (rs2228570), and TaqI (rs731236). METHODS: PubMed, Web of Science, CBM (Chinese Biomedical Database), CNKI (China National Knowledge Infrastructure), and Wanfang (Chinese) database were searched for relevant studies...
August 23, 2016: Pediatric Pulmonology
Juliana Maia, Andreia Soares da Silva, Rodrigo Feliciano do Carmo, Taciana Furtado de Mendonça, Luiz Henrique Maciel Griz, Patricia Moura, Francisco Bandeira
INTRODUCTION: Since there is evidence of the action of vitamin D as a modulator of insulin release and atherosclerosis, it may well be that the vitamin D receptor polymorphisms are associated with diabetes and its chronic complications. AIMS: To examine the associations between vitamin D receptor polymorphisms (FokI and TaqI) and Type 2 diabetes (T2DM) and its associated chronic complications in postmenopausal women. METHODS: This cross-sectional study analyzed 100 postmenopausal women with T2DM (mean age 65...
2016: Application of Clinical Genetics
Wael Abdel Halim Hegazy, Mahmoud Youns
Cancer is thought to be a direct result of transcriptional misregulation. Broad analysis of transcriptional regulatory elements in healthy and cancer cells is needed to understand cancer development. Nucleases regulatory domains are recruited to bind and manipulate a specific genomic locus with high efficacy and specificity. TALENs (transcription activatorlike effector nuclease) fused to endonuclease FokI have been used widely to target specific sequences to edit several genes in healthy and cancer cells. This approach is promising to target specific cancer genes and for this purpose it is needed to pack such TALENs into viral vectors...
2016: Asian Pacific Journal of Cancer Prevention: APJCP
Soranun Chantarangsu, Thanyachai Sura, Sanutm Mongkornkarn, Kobkiat Donsakul, Kitti Torrungruang
BACKGROUND: Polymorphisms of the vitamin D receptor (VDR) gene have been implicated in the susceptibility to infections and bone-related diseases. However, their relationship with periodontal disease remains unclear. This cross-sectional study investigated whether the susceptibility to chronic periodontitis (CP) in a Thai population is associated with VDR polymorphisms. METHODS: Genomic DNA was obtained from 1,460 subjects, aged 39-66. Genotyping of VDR polymorphisms (FokI, BsmI, ApaI, and TaqI) was performed using real-time polymerase chain reaction...
August 4, 2016: Journal of Periodontology
Dilsah Cogulu, Huseyin Onay, Yasemin Ozdemir, Gulcin Itirli Aslan, Ferda Ozkinay, Cemal Eronat
OBJECTIVE: To determine the association between the ApaI, FokI, Cdx2 and TaqI polymorphisms of vitamin D receptor (VDR) gene in caries-active (high-moderate) and caries-free children. STUDY DESIGN: A hundred and fifty children (75 males, 75 females, mean age: 10.19 ± 1.61 years) were included in the study. The subjects were divided into three groups as high caries risk group (DMFT, dft>4)(n=55), moderate caries risk group (DMFT, dft=1-4)(n=57) and caries-free group (n=38)...
2016: Journal of Clinical Pediatric Dentistry
Bedia Guleryuz, Fulya Akin, Melek T Ata, Mukaddes Mergen Dalyanoglu, Sebahat Turgut
BACKGROUND: Hashimoto's thyroiditis (HT) is a common autoimmune disease. Vitamin D is an important regulator of immune system. It has been shown in several studies that vitamin D prevents the development of lots of autoimmune diseases. There are some studies that prove vitamin D receptor (VDR) gene polymorphism increases the risk of Hashimoto's thyroiditis. In this study, we aimed to investigate the association between HT and level of 25(OH)D3, IL-2, IL-4, IL-5, TNF-α and IFN-γ and VDR FokI and TaqI gene polymorphism...
July 27, 2016: Endocrine, Metabolic & Immune Disorders Drug Targets
Fei Yu, Ling Ling Cui, Xing Li, Chong Jian Wang, Yue Ba, Ling Wang, Jing Li, Chao Li, Li Ping Dai, Wen Jie Li
BACKGROUND AND OBJECTIVES: Vitamin D receptor (VDR) genetic polymorphisms are considered to be associated with type 2 diabetes mellitus (T2DM), but this is inconclusive. The aim of this study is to quantify the association between polymorphisms of BsmI and FokI in the VDR gene and T2DM risk through literature review. METHODS AND STUDY DESIGN: Original articles published from 1999 to June 2014 were discovered through PubMed, ISI Web of Science, China National Knowledge Infrastructure, Chinese Wanfang Database, and the Chinese Biomedical Literature Database...
2016: Asia Pacific Journal of Clinical Nutrition
Rasoul Abdollahzadeh, Mahsa Sobhani Fard, Farideh Rahmani, Kaveh Moloudi, Behrooz Sadeghi Kalani, Asaad Azarnezhad
Multiple sclerosis (MS) is a chronic inflammatory, demyelinating, and neurodegenerative disease of the central nervous system (CNS) with a complex etiology. Given the Vitamin D receptor (VDR) gene, it is considered an outstanding risk component associated with MS. The aim of the present study has been to explore and emphasize the role of ApaI, BsmI, TaqI and FokI polymorphisms of VDR gene in susceptibility to MS in an Iranian case-control population including 160 patients and 150 healthy controls. All cases were clinically diagnosed with relapsing-remitting (RR) form, and the controls were age, gender, and race matched which were completely in agreement with the case group...
August 15, 2016: Journal of the Neurological Sciences
Shereen Sobeih, Heba M Mashaly, Heba Gawdat, Khalda Amr, Mahmoud Fawzy Abdel Hamid, Eman Shaalan
BACKGROUND: Vitiligo is an autoimmune depigmentation disorder. Polymorphisms in the vitamin D receptor (VDR) have been found to be associated with vitiligo. OBJECTIVES: To evaluate the potential association between VDR gene polymorphisms (ApaI, TaqI, and FokI) and vitiligo susceptibility, and to detect if there is correlation between serum 25-hydroxyvitamin D [25(OH)D] levels and vitiligo and between VDR gene polymorphisms and 25(OH)D levels in vitiligo. MATERIALS AND METHODS: Seventy-five patients with vitiligo and 75 age and sex-matched controls were subjected to detailed history taking and dermatological examination to determine the extent and clinical type of vitiligo...
July 15, 2016: International Journal of Dermatology
Seo Young Kang, Suyeon Park, Eungseok Oh, Jinse Park, Jinyoung Youn, Ji Sun Kim, Jeong-Uk Kim, Wooyoung Jang
Recently, the effect of genetic variants in the Vitamin D receptor (VDR) gene on Parkinson's disease (PD) has gained interest. However, the precise relationship between VDR polymorphisms and PD remains unclear. In Korea, one study reported an association between VDR gene polymorphisms and PD. However, this study was conducted with a small sample size, and only the Bsml locus was evaluated. Therefore, further investigations about the relationship between VDR polymorphisms and PD are necessary in a Korean population...
August 15, 2016: Neuroscience Letters
Hoda Rasheed, Rehab A Hegazy, Heba I Gawdat, Dina A Mehaney, Marwa M Kamel, Marwa M Fawzy, Mohammed M Nooh, Hebatallah A Darwish
BACKGROUND: Vitamin D has been considered a key player in various malignancies including cutaneous cancers. To date, mycosis fungoides (MF) has been the least studied in relation to vitamin D. Furthermore, the vitamin D receptor (VDR) single nucleotide polymorphisms (SNPs) have not been tackled before in the context of MF, despite their incrimination in numerous diseases. AIM OF STUDY: To assess the role of vitamin D in MF by measuring its serum level, and studying VDR SNPs (TaqI, BsmI, FokI) in different stages of MF...
2016: PloS One
Nan Jiang, Xing-Qi Zhao, Cheng-He Qin, Yan-Jun Hu, Lei Wang, Guo-Ping Xie, Sheng-Nan Wang, Li-Guang Chen, Bin Yu
Previous studies have indicated that vitamin D receptor (VDR) TaqI, BsmI, FokI and ApaI gene polymorphisms are associated with the risk of several inflammatory diseases. However, potential association of the VDR gene polymorphisms with susceptibility to extremity chronic osteomyelitis remains unclear. The present study aimed to investigate link between VDR gene polymorphisms and the risk of extremity chronic osteomyelitis in Chinese population. A total of 233 patients with chronic osteomyelitis and 200 healthy controls were genotyped for the 4 single-nucleotide polymorphisms (SNPs) (TaqI, BsmI, FokI and ApaI) in VDR gene using the SNaPshot genotyping method...
August 2016: Injury
Khayal A Alkhayal, Zainab H Awadalia, Mansoor-Ali Vaali-Mohammed, Omar A Al Obeed, Alanoud Al Wesaimer, Rabih Halwani, Ahmed M Zubaidi, Zahid Khan, Maha-Hamadien Abdulla
BACKGROUND: Vitamin D, causally implicated in bone diseases and human malignancies, exerts its effects through binding to the vitamin D receptor (VDR). VDR is a transcription factor modulating the expression of several genes in different pathways. Genetic variants in the VDR gene have been associated with several cancers in different population including colorectal cancer. OBJECTIVE: To assess the association of VDR gene polymorphisms in relation with colorectal cancer (CRC) in a Saudi population...
2016: PloS One
Sha Liao, Jun He, Xiaoxia Li, Honexia Xu, Xiuying Liu, Yi Zhao, Yuhong Zhang
OBJECTIVE: To explore the association between the vitamin D receptor (VDR) gene polymorphisms and HOMA index in prediabetes. METHODS: On the basis of a cross-sectional study which was conducted in Ningxia during 2008-2012, 339 controls and 468 subjects with prediabetes were selected according to ADA diabetes diagnosis standards. Anthropometric data and blood samples were collected in the field investigation. Blood biochemistry analyses and insulin determination were carried out in the laboratory...
March 2016: Wei Sheng Yan Jiu, Journal of Hygiene Research
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