RNAi-mediated Reduction of a Major Ruminant Specific Milk Allergen Using a Transgenic Mouse Model

Milk from dairy cows is an important human food. It contains the protein beta-lactoglobulin (BLG) which is not present in human milk and thought to be a major allergen in the milk of cows. RNA interference (RNAi), mediated by sequence-specific micro RNAs (miRNAs), provides a new molecular tool to reduce the levels of undesirable proteins. In this study, I have screened ten miRNAs targeting the BLG mRNA for their potential to knockdown BLG expression using a cell-based in vitro assay. I identified four miRNAs which showed substantial bovine BLG (bBLG) (78-97%) and ovine BLG (oBLG) (33-94%) knockdown activity in vitro. Tandem miRNA constructs with combinations of these four single miRNAs did not result in significantly increased knockdown efficiency compared to the respective single miRNA constructs in the in vitro assay (P>0.05). Because targeting of two different regions has the potential for greater knockdown efficiency in vivo, I chose a tandem construct which achieved in vitro up to 99% bBLG knockdown as compared to 92% and 74% of bBLG knockdown by the single miRNAs. The tandem construct also showed 90% of oBLG knockdown in comparison to 92% and 69% of oBLG knockdown by the single miRNAs in the in vitro assay which made it an ideal candidate for the subsequent evaluation in transgenic (Tg) mice. For the in vivo studies, the cytomegalovirus (CMV) promoter of the selected miRNA constructs was replaced with the promoter of the mouse milk protein gene for whey acid protein (WAP) which directs the expression of the BLG-specific miRNAs to the lactating mammary gland. The Tg mouse lines generated with the tissue-specific tandem construct and the single miRNAs used for the tandem miRNA construct were then crossed with an oBLG or bBLG expressing Tg mouse line to generate double transgenic (miRNA-BLG) mice for assessing BLG knockdown in vivo. Analysis of the milk proteins demonstrated that the tandem miRNA reduced the levels of oBLG and bBLG in milk from the miRNA-BLG mice up to 96% and 46%, respectively. This study validates the mammary gland specific expression of miRNAs as a promising approach to knockdown allergenic proteins in milk

Prostate Cancer: Is It a Battle Lost to Age?

Age is often considered an important non-modifiable risk factor for a number of diseases, including prostate cancer. Some prominent risk factors of prostate cancer include familial history, ethnicity and age. In this review, various genetic and physiological characteristics affected due to advancing age will be analysed and correlated with their direct effect on prostate cancer

Effect of ageing and single nucleotide polymorphisms associated with the risk of aggressive prostate cancer in a New Zealand population

Prostate cancer is one of the most significant male health concerns worldwide, and various researchers carrying out molecular diagnostics have indicated that genetic interactions with biological and behavioral factors play an important role in the overall risk and prognosis of this disease. Single nucleotide polymorphisms are increasingly becoming strong biomarker candidates to identify the susceptibility of individuals to prostate cancer. We carried out risk association of different stages of prostate cancer to a number of single nucleotide polymorphisms to identify the susceptible alleles in a New Zealand population and checked the interaction with environmental factors as well. We identified a number of single nucleotide polymorphisms to have associations specifically to the risk of prostate cancer and aggressiveness of the disease, and also certain single nucleotide polymorphisms to be vulnerable to the reported behavioral factors. We have addressed “special” environmental conditions prevalent in New Zealand, which can be used as a model for a bigger worldwide study

Environmental factors and risk of aggressive prostate cancer among a population of New Zealand men - a genotypic approach

Prostate cancer is one of the most significant health concerns for men worldwide. Numerous researchers carrying out molecular diagnostics have indicated that genetic interactions with biological and behavioral factors play an important role in the overall risk and prognosis of this disease. Single nucleotide polymorphisms (SNPs) are increasingly becoming strong biomarker candidates to identify susceptibility to prostate cancer. We carried out a gene × environment interaction analysis linked to aggressive and non-aggressive prostate cancer (PCa) with a number of SNPs. By using this method, we identified the susceptible alleles in a New Zealand population, and examined the interaction with environmental factors. We have identified a number of SNPs that have risk associations both with and without environmental interaction. The results indicate that certain SNPs are associated with disease vulnerability based on behavioral factors. The list of genes with SNPs identified as being associated with the risk of PCa in a New Zealand population is provided in the graphical abstrac

SNP-SNP interactions as risk factors for aggressive prostate cancer [version 1; referees: 2 approved]

Prostate cancer (PCa) is one of the most significant male health concerns worldwide. Single nucleotide polymorphisms (SNPs) are becoming increasingly strong candidate biomarkers for identifying susceptibility to PCa. We identified a number of SNPs reported in genome-wide association analyses (GWAS) as risk factors for aggressive PCa in various European populations, and then defined SNP-SNP interactions, using PLINK software, with nucleic acid samples from a New Zealand cohort. We used this approach to find a gene x environment marker for aggressive PCa, as although statistically gene x environment interactions can be adjusted for, it is highly impossible in practicality, and thus must be incorporated in the search for a reliable biomarker for PCa. We found two intronic SNPs statistically significantly interacting with each other as a risk for aggressive prostate cancer on being compared to healthy controls in a New Zealand population

Correction: Vaidyanathan et al. Are We Eating Our Way to Prostate Cancer—A Hypothesis Based on the Evolution, Bioaccumulation, and Interspecific Transfer of miR-150. Non-Coding RNA 2016, 2, 2.

The authors wish to make the following correction to this paper [1].[...