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"James, Anthony"
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Bombyx mori P-element Somatic Inhibitor (BmPSI) Is a Key Auxiliary Factor for Silkworm Male Sex Determination
Manipulation of sex determination pathways in insects provides the basis for a wide spectrum of strategies to benefit agriculture and public health. Furthermore, insects display a remarkable diversity in the genetic pathways that lead to sex differentiation. The silkworm, Bombyx mori, has been cultivated by humans as a beneficial insect for over two millennia, and more recently as a model system for studying lepidopteran genetics and development. Previous studies have identified the B. mori Fem piRNA as the primary female determining factor and BmMasc as its downstream target, while the genetic scenario for male sex determination was still unclear. In the current study, we exploite the transgenic CRISPR/Cas9 system to generate a comprehensive set of knockout mutations in genes BmSxl, Bmtra2, BmImp, BmImpM, BmPSI and BmMasc, to investigate their roles in silkworm sex determination. Absence of Bmtra2 results in the complete depletion of Bmdsx transcripts, which is the conserved downstream factor in the sex determination pathway, and induces embryonic lethality. Loss of BmImp or BmImpM function does not affect the sexual differentiation. Mutations in BmPSI and BmMasc genes affect the splicing of Bmdsx and the female reproductive apparatus appeared in the male external genital. Intriguingly, we identify that BmPSI regulates expression of BmMasc, BmImpM and Bmdsx, supporting the conclusion that it acts as a key auxiliary factor in silkworm male sex determination.
Journal Article
Transgenic Cavendish bananas with resistance to Fusarium wilt tropical race 4
Banana (
Musa
spp.) is a staple food for more than 400 million people. Over 40% of world production and virtually all the export trade is based on Cavendish banana. However, Cavendish banana is under threat from a virulent fungus,
Fusarium oxysporum
f. sp.
cubense
tropical race 4 (TR4) for which no acceptable resistant replacement has been identified. Here we report the identification of transgenic Cavendish with resistance to TR4. In our 3-year field trial, two lines of transgenic Cavendish, one transformed with
RGA2
, a gene isolated from a TR4-resistant diploid banana, and the other with a nematode-derived gene,
Ced9
, remain disease free. Transgene expression in the RGA2 lines is strongly correlated with resistance. Endogenous RGA2 homologs are also present in Cavendish but are expressed tenfold lower than that in our most resistant transgenic line. The expression of these homologs can potentially be elevated through gene editing, to provide non-transgenic resistance.
The newly recognized Fusarium wilt pathogen tropical race 4 is threatening worldwide banana production. Here, the authors transform Cavendish bananas with a resistance gene,
RGA2
, from diploid banana or a nematode-derived gene,
Ced9
, and confer resistance to natural infection under field conditions.
Journal Article
Single-shot on-chip spectral sensors based on photonic crystal slabs
Miniaturized spectrometers have significant potential for portable applications such as consumer electronics, health care, and manufacturing. These applications demand low cost and high spectral resolution, and are best enabled by single-shot free-space-coupled spectrometers that also have sufficient spatial resolution. Here, we demonstrate an on-chip spectrometer that can satisfy all of these requirements. Our device uses arrays of photodetectors, each of which has a unique responsivity with rich spectral features. These responsivities are created by complex optical interference in photonic-crystal slabs positioned immediately on top of the photodetector pixels. The spectrometer is completely complementary metal–oxide–semiconductor (CMOS) compatible and can be mass produced at low cost.
Miniaturized components have significant potential for portable applications. Here, the authors demonstrate a spectrometer and hyperspectral imager based on photonic-crystal slabs which can enable single-shot imaging on a compact chip.
Journal Article
Will Hong Kong ever have an ethical research system?
Hong Kong universities have animal ethics committees but standards of ethical review among the various committees are highly variable in their effectiveness. Since 1999, the DoH has become incrementally more proactive as pressure has been applied by stakeholders in animal welfare. [...]my recent personal experience has revealed that self-regulated ethics committees still lack effectiveness because I have observed unacceptable experimental techniques, for example, failure to use analgesia, failure to reduce animal numbers or consider replacements to animal models and numerous non-compliance issues by researchers with their DoH licences and ethics committee conditions. Self-regulation alone is inadequate. ◾ No pressure is exerted from stakeholders (eg, funders and publishers) on researchers to comply with international standards for the ethical use of animals in research. ◾ There is no commitment from senior levels of university management to ensure researchers adhere to the highest ethical standards when using animals in research. ◾ And that there is no effective animal ethical review process.
Journal Article
Highly efficient Cas9-mediated gene drive for population modification of the malaria vector mosquito Anopheles stephensi
Malaria continues to impose enormous health and economic burdens on the developing world. Novel technologies proposed to reduce the impact of the disease include the introgression of parasite-resistance genes into mosquito populations, thereby modifying the ability of the vector to transmit the pathogens. Such genes have been developed for the human malaria parasite Plasmodium falciparum . Here we provide evidence for a highly efficient gene-drive system that can spread these antimalarial genes into a target vector population. This system exploits the nuclease activity and target-site specificity of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system, which, when restricted to the germ line, copies a genetic element from one chromosome to its homolog with ≥98% efficiency while maintaining the transcriptional activity of the genes being introgressed. Genetic engineering technologies can be used both to create transgenic mosquitoes carrying antipathogen effector genes targeting human malaria parasites and to generate gene-drive systems capable of introgressing the genes throughout wild vector populations. We developed a highly effective autonomous Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein 9 (Cas9)-mediated gene-drive system in the Asian malaria vector Anopheles stephensi , adapted from the mutagenic chain reaction (MCR). This specific system results in progeny of males and females derived from transgenic males exhibiting a high frequency of germ-line gene conversion consistent with homology-directed repair (HDR). This system copies an ∼17-kb construct from its site of insertion to its homologous chromosome in a faithful, site-specific manner. Dual anti- Plasmodium falciparum effector genes, a marker gene, and the autonomous gene-drive components are introgressed into ∼99.5% of the progeny following outcrosses of transgenic lines to wild-type mosquitoes. The effector genes remain transcriptionally inducible upon blood feeding. In contrast to the efficient conversion in individuals expressing Cas9 only in the germ line, males and females derived from transgenic females, which are expected to have drive component molecules in the egg, produce progeny with a high frequency of mutations in the targeted genome sequence, resulting in near-Mendelian inheritance ratios of the transgene. Such mutant alleles result presumably from nonhomologous end-joining (NHEJ) events before the segregation of somatic and germ-line lineages early in development. These data support the design of this system to be active strictly within the germ line. Strains based on this technology could sustain control and elimination as part of the malaria eradication agenda.
Journal Article