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

authors: Jabed, Md Anower

subjects: BLG, miRNA, RNAi, Transgenic mouse, Gene knockdown

abstract: 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.

text: 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.

year: 2012

title: Introduction: Religious rupture and revival in the Pacific

authors: Macdonald, Fraser

subjects: Science & Technology

abstract: This special issue arose from a panel, “Political and Religious Conversions in the Pacific”, convened by Fraser Macdonald and Michael Goldsmith at the 2017 Australian Anthropological Society (AAS) annual conference held at the University of Adelaide. The papers carried over from that session into this special issue more directly address the question of religious conversion, marshalling evidence from a wide range of ethnographic contexts throughout the Pacific to shed light on the ramifications of conversion to various brands of Christianity. The issue collectively points up a number of critical issues now gaining deeper appreciation within anthropological writing on Pacific Christianity, including the centrality of indigenous agency to processes of change; the importance of charismatic leaders who initiate and orchestrate popular movements; the intrinsically political character of Christianity in the Pacific, including its inextricable historical relationship with colonialism; and, finally, the strongly regional dimensions of Christianity, whereby particular societies are embedded in and contribute to broader religious communities that transcend the local.

text: Introduction: Religious rupture and revival in the Pacific This special issue arose from a panel, “Political and Religious Conversions in the Pacific”, convened by Fraser Macdonald and Michael Goldsmith at the 2017 Australian Anthropological Society (AAS) annual conference held at the University of Adelaide. The papers carried over from that session into this special issue more directly address the question of religious conversion, marshalling evidence from a wide range of ethnographic contexts throughout the Pacific to shed light on the ramifications of conversion to various brands of Christianity. The issue collectively points up a number of critical issues now gaining deeper appreciation within anthropological writing on Pacific Christianity, including the centrality of indigenous agency to processes of change; the importance of charismatic leaders who initiate and orchestrate popular movements; the intrinsically political character of Christianity in the Pacific, including its inextricable historical relationship with colonialism; and, finally, the strongly regional dimensions of Christianity, whereby particular societies are embedded in and contribute to broader religious communities that transcend the local.

year: 2019-12-01

title: Kūkupa, koro, and kai: The use of Māori vocabulary items in New Zealand English children's picture books

authors: Daly, Nicola

subjects: New Zealand

abstract: When a linguistic form from one language is used in another language, such words are known as borrowings or loan words (Crystal 2003: 56). The English language is renowned for its large capacity for borrowing and it has been suggested that the growth in internationalism in recent times has led to people seeking new words to indicate their local identity (Crystal 1995; Crystal 2003). Certainly this is the case of New Zealand English, the most distinctive aspect of which is borrowings from te reo Maori (Deverson 1991). In 1984 Deverson estimated that most New Zealanders have a passive knowledge of at least 40-50 borrowed Maori loan words (Deverson 1984). This figure has been recently revised by Macalister to 70-80 such words (20,-,63.). A study of the frequency of Maori loan words in New Zealand English in New Zealand School Journals of the 1960s and 1990s showed an incidence of around 6 words per 1,000 (Macalister 1999). Kennedy and Yamazaki (1999) also found borrowed Maori words at a rate of 6 per thousand words. Macalister (2006b) has examined the use of Maori loan words in New Zealand English across a 150 year period from 1850-2000. He examined a corpus of a little under five and a half million words from three sources: Newspapers, parliamentary debates and School Journals. Across the three sources there was an increase from 3.29 words per 1,000 in 1850 to 8.8 per thousand in 2000. Macalister lists the reasons for this change as including urbanisation of the Maori population between 1945 and 1975, which created more contact; the changing status of the Maori language with the kOhanga reo movement; the establishment of the Maori Language Commission (Te Taura Whiri i te Reo Maori) and legislation of Maori as an official language of New Zealand in 1987.

text: Kūkupa, koro, and kai: The use of Māori vocabulary items in New Zealand English children's picture books When a linguistic form from one language is used in another language, such words are known as borrowings or loan words (Crystal 2003: 56). The English language is renowned for its large capacity for borrowing and it has been suggested that the growth in internationalism in recent times has led to people seeking new words to indicate their local identity (Crystal 1995; Crystal 2003). Certainly this is the case of New Zealand English, the most distinctive aspect of which is borrowings from te reo Maori (Deverson 1991). In 1984 Deverson estimated that most New Zealanders have a passive knowledge of at least 40-50 borrowed Maori loan words (Deverson 1984). This figure has been recently revised by Macalister to 70-80 such words (20,-,63.). A study of the frequency of Maori loan words in New Zealand English in New Zealand School Journals of the 1960s and 1990s showed an incidence of around 6 words per 1,000 (Macalister 1999). Kennedy and Yamazaki (1999) also found borrowed Maori words at a rate of 6 per thousand words. Macalister (2006b) has examined the use of Maori loan words in New Zealand English across a 150 year period from 1850-2000. He examined a corpus of a little under five and a half million words from three sources: Newspapers, parliamentary debates and School Journals. Across the three sources there was an increase from 3.29 words per 1,000 in 1850 to 8.8 per thousand in 2000. Macalister lists the reasons for this change as including urbanisation of the Maori population between 1945 and 1975, which created more contact; the changing status of the Maori language with the kOhanga reo movement; the establishment of the Maori Language Commission (Te Taura Whiri i te Reo Maori) and legislation of Maori as an official language of New Zealand in 1987.

year: 2007