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Article ; Online: The Transcriptome of Chicken Migratory Primordial Germ Cells Reveals Intrinsic Sex Differences and Expression of Hallmark Germ Cell Genes.

Doddamani, Dadakhalandar / Woodcock, Mark / Taylor, Lorna / Nandi, Sunil / McTeir, Lynn / Davey, Megan G / Smith, Jacqueline / McGrew, Mike J

Cells

2023 Volume 12, Issue 8

Abstract: Primordial germ cells (PGCs) are germline-restricted embryonic cells that form the functional gametes of the adult animal. The use of avian PGCs in biobanking and producing genetically modified birds has driven research on the in vitro propagation and ... ...

Abstract	Primordial germ cells (PGCs) are germline-restricted embryonic cells that form the functional gametes of the adult animal. The use of avian PGCs in biobanking and producing genetically modified birds has driven research on the in vitro propagation and manipulation of these embryonic cells. In avian species, PGCs are hypothesized to be sexually undetermined at an early embryonic stage and undergo differentiation into an oocyte or spermatogonial fate dictated by extrinsic factors present in the gonad. However, chicken male and female PGCs require different culture conditions, suggesting that there are sex-specific differences, even at early stages. To understand potential differences between male and female chicken PGCs during migratory stages, we studied the transcriptomes of circulatory stage male and female PGCs propagated in a serum-free medium. We found that in vitro cultured PGCs were transcriptionally similar to their in ovo counterparts, with differences in cell proliferation pathways. Our analysis also revealed sex-specific transcriptome differences between male and female cultured PGCs, with notable differences in Smad7 and NCAM2 expression. A comparison of chicken PGCs with pluripotent and somatic cell types identified a set of genes that are exclusive to germ cells, enriched in the germplasm, and associated with germ cell development.
MeSH term(s)	Animals ; Female ; Male ; Chickens/genetics ; Transcriptome/genetics ; Sex Characteristics ; Biological Specimen Banks ; Germ Cells/metabolism
Language	English
Publishing date	2023-04-13
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12081151
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Antibody Responses to SARS-CoV-2 Infection-Comparative Determination of Seroprevalence in Two High-Throughput Assays versus a Sensitive Spike Protein ELISA.

Mohanraj, Dinesh / Bicknell, Kelly / Bhole, Malini / Webber, Caroline / Taylor, Lorna / Whitelegg, Alison

Vaccines

2021 Volume 9, Issue 11

Abstract: Robust assay development for SARS-CoV-2 serological testing requires assessment of asymptomatic and non-hospitalised individuals to determine if assays are sensitive to mild antibody responses. Our study evaluated the performance characteristics of two ... ...

Abstract	Robust assay development for SARS-CoV-2 serological testing requires assessment of asymptomatic and non-hospitalised individuals to determine if assays are sensitive to mild antibody responses. Our study evaluated the performance characteristics of two high-throughput SARS-CoV-2 IgG nucleocapsid assays (Abbott Architect and Roche) and The Binding Site (TBS) Anti-Spike IgG/A/M ELISA kit in samples from healthcare workers (HCWs). The 252 samples were collected from multi-site NHS trusts and analysed for SARS-CoV-2 serology. Assay performance was evaluated between these three platforms and ROC curves were used to redefine the Abbott threshold. Concordance between Abbott and TBS was 66%. Any discrepant results were analysed using Roche, which showed 100% concordance with TBS. Analysis conducted in HCWs within 58 days post-PCR result demonstrated 100% sensitivity for both Abbott and Roche. Longitudinal analysis for >100 days post-PCR led to sensitivity of 77.2% and 100% for Abbott and Roche, respectively. A redefined Abbott threshold (0.64) increased sensitivity to 90%, producing results comparable to TBS and Roche. The manufacturer's threshold set by Abbott contributes to lower sensitivity and elevated false-negative occurrences. Abbott performance improved upon re-optimisation of the cut-off threshold. Our findings provided evidence that TBS can be used as bespoke alternative for SARS-CoV-2 serology analysis where high-throughput platforms are not feasible on site.
Language	English
Publishing date	2021-11-11
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines9111310
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Article ; Online: Direct allele introgression into pure chicken breeds using Sire Dam Surrogate (SDS) mating.

Ballantyne, Maeve / Woodcock, Mark / Doddamani, Dadakhalandar / Hu, Tuanjun / Taylor, Lorna / Hawken, Rachel J / McGrew, Mike J

Nature communications

2021 Volume 12, Issue 1, Page(s) 659

Abstract: Poultry is the most abundant livestock species with over 60 billion chickens raised globally per year. The majority of chicken are produced from commercial flocks, however many indigenous chicken breeds play an important role in rural economies as they ... ...

Abstract	Poultry is the most abundant livestock species with over 60 billion chickens raised globally per year. The majority of chicken are produced from commercial flocks, however many indigenous chicken breeds play an important role in rural economies as they are well adapted to local environmental and scavenging conditions. The ability to make precise genetic changes in chicken will permit the validation of genetic variants responsible for climate adaptation and disease resilience, and the transfer of beneficial alleles between breeds. Here, we generate a novel inducibly sterile surrogate host chicken. Introducing donor genome edited primordial germ cells into the sterile male and female host embryos produces adult chicken carrying only exogenous germ cells. Subsequent direct mating of the surrogate hosts, Sire Dam Surrogate (SDS) mating, recreates the donor chicken breed carrying the edited allele in a single generation. We demonstrate the introgression and validation of two feather trait alleles, Dominant white and Frizzle into two pure chicken breeds using the SDS surrogate hosts.
MeSH term(s)	Alleles ; Animals ; Breeding/methods ; Chickens/genetics ; Feathers ; Female ; Germ Cells/metabolism ; Infertility/genetics ; Male ; Phenotype ; Reproducibility of Results ; Reproduction/genetics
Language	English
Publishing date	2021-01-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-020-20812-x
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Comparative assessment of SARS-CoV-2 serology in healthcare workers with Abbott Architect, Roche Elecsys and The Binding site ELISA immunoassays.

Mohanraj, Dinesh / Whitelegg, Alison / Bicknell, Kelly / Bhole, Malini / Webber, Caroline / Taylor, Lorna

medRxiv

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology testing is key for assessing seroprevalence and antibody response post-vaccination in immunocompromised patients. Here we performed a comparison between two high-throughput ... ...

Abstract	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology testing is key for assessing seroprevalence and antibody response post-vaccination in immunocompromised patients. Here we performed a comparison between two high-throughput nucleocapsid assays (Abbott SARS-CoV-2 IgG and Roche Elecsys Anti-SARS-CoV-2) and The Binding Site (TBS) anti-Spike IgG/A/M- SARS-CoV-2 ELISA kit. 236 samples were collected across 2 sites, Portsmouth Hospital University NHS Trust (PHU) and The Dudley Group NHS Trust. We derived concordance, agreement and assay performance as well as using receiver operating characteristic (ROC) curves to redefine the assay threshold of the Abbott assay. Result concordance between the Abbott and TBS was 66%. Discrepant samples were analysed using the Roche assay which showed 100% agreement with the TBS assay. In samples analysed >58 days post-PCR, the sensitivity of Abbott and Roche was 100%. In samples analysed >100 days post-PCR the sensitivity of the Abbott assay dropped to 77.2% but remained at 100% for the Roche assay. A redefined Abbott threshold of 0.64 increased the sensitivity to 90% giving results similar to the Roche and TBS assays. In conclusion, this study demonstrated Abbott assay had a lower sensitivity in comparison to TBS and Roche. This study established TBS can be implemented as a viable alternative for SARS-CoV-2 serology testing where high-throughput assays are not available on site. Furthermore, anti-spike assays, such as TBS, could be used to monitor vaccination responses to deduce SARS-CoV-2 population-immunity. Further optimization studies are required to evaluate the performance characteristics of these assays which could facilitate widescale sero-epidemiological surveillance.
Keywords	covid19
Language	English
Publishing date	2021-03-24
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2021.03.19.21253518
Database	COVID19

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Article ; Online: A low-tech, cost-effective and efficient method for safeguarding genetic diversity by direct cryopreservation of poultry embryonic reproductive cells.

Hu, Tuanjun / Taylor, Lorna / Sherman, Adrian / Keambou Tiambo, Christian / Kemp, Steven J / Whitelaw, Bruce / Hawken, Rachel J / Djikeng, Appolinaire / McGrew, Michael J

eLife

2022 Volume 11

Abstract: Chickens are an important resource for smallholder farmers who raise locally adapted, genetically distinct breeds for eggs and meat. The development of efficient reproductive technologies to conserve and regenerate chicken breeds safeguards existing ... ...

Abstract	Chickens are an important resource for smallholder farmers who raise locally adapted, genetically distinct breeds for eggs and meat. The development of efficient reproductive technologies to conserve and regenerate chicken breeds safeguards existing biodiversity and secures poultry genetic resources for climate resilience, biosecurity, and future food production. The majority of the over 1600 breeds of chicken are raised in low and lower to middle income countries under resource-limited, small-scale production systems, which necessitates a low-tech, cost-effective means of conserving diversity is needed. Here, we validate a simple biobanking technique using cryopreserved embryonic chicken gonads. The gonads are quickly isolated, visually sexed, pooled by sex, and cryopreserved. Subsequently, the stored material is thawed and dissociated before injection into sterile host chicken embryos. By using pooled GFP and RFP-labelled donor gonadal cells and Sire Dam Surrogate mating, we demonstrate that chicks deriving entirely from male and female donor germ cells are hatched. This technology will enable ongoing efforts to conserve chicken genetic diversity for both commercial and smallholder farmers, and to preserve existing genetic resources at poultry research facilities.
MeSH term(s)	Animals ; Biological Specimen Banks ; Breeding/methods ; Chickens/genetics ; Chickens/physiology ; Cost-Benefit Analysis ; Cryopreservation/veterinary ; Female ; Genetic Variation ; Germ Cells/cytology ; Infertility/veterinary ; Male
Language	English
Publishing date	2022-01-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.74036
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: High fidelity CRISPR/Cas9 increases precise monoallelic and biallelic editing events in primordial germ cells.

Idoko-Akoh, Alewo / Taylor, Lorna / Sang, Helen M / McGrew, Michael J

Scientific reports

2018 Volume 8, Issue 1, Page(s) 15126

Abstract: Primordial germ cells (PGCs), the embryonic precursors of the sperm and egg, are used for the introduction of genetic modifications into avian genome. Introduction of small defined sequences using genome editing has not been demonstrated in bird species. ...

Abstract	Primordial germ cells (PGCs), the embryonic precursors of the sperm and egg, are used for the introduction of genetic modifications into avian genome. Introduction of small defined sequences using genome editing has not been demonstrated in bird species. Here, we compared oligonucleotide-mediated HDR using wild type SpCas9 (SpCas9-WT) and high fidelity SpCas9-HF1 in PGCs and show that many loci in chicken PGCs can be precise edited using donors containing CRISPR/Cas9-blocking mutations positioned in the protospacer adjacent motif (PAM). However, targeting was more efficient using SpCas9-HF1 when mutations were introduced only into the gRNA target sequence. We subsequently employed an eGFP-to-BFP conversion assay, to directly compare HDR mediated by SpCas9-WT and SpCas9-HF1 and discovered that SpCas9-HF1 increases HDR while reducing INDEL formation. Furthermore, SpCas9-HF1 increases the frequency of single allele editing in comparison to SpCas9-WT. We used SpCas9-HF1 to demonstrate the introduction of monoallelic and biallelic point mutations into the FGF20 gene and generate clonal populations of edited PGCs with defined homozygous and heterozygous genotypes. Our results demonstrate the use of oligonucleotide donors and high fidelity CRISPR/Cas9 variants to perform precise genome editing with high efficiency in PGCs.
MeSH term(s)	Alleles ; Animals ; Base Sequence ; Binding Sites ; CRISPR-Cas Systems ; Chickens ; Gene Editing ; Gene Order ; Genetic Vectors/genetics ; Germ Cells/cytology ; Germ Cells/metabolism ; Heterozygote ; INDEL Mutation ; Mutation ; Protein Binding ; Sequence Analysis, DNA
Language	English
Publishing date	2018-10-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-018-33244-x
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Learning Style Preferences of Practicing Nurses.

Mangold, Kara / Kunze, Katie L / Quinonez, Michelle M / Taylor, Lorna M / Tenison, Ashley J

Journal for nurses in professional development

2018 Volume 34, Issue 4, Page(s) 212–218

Abstract: Nursing professional development practitioners are encouraged to consider incorporating preferred learning styles into professional development programs. However, conclusive evidence about preferred learning styles does not exist. This study describes ... ...

Abstract	Nursing professional development practitioners are encouraged to consider incorporating preferred learning styles into professional development programs. However, conclusive evidence about preferred learning styles does not exist. This study describes the preferred learning styles of nursing staff. Results showed that learning style preferences existed and were correlated with satisfaction, years of experience, and gender. The results can be used to plan and deliver professional development opportunities that are engaging and promote retention of learning.
MeSH term(s)	Female ; Humans ; Learning ; Male ; Models, Educational ; Nursing Education Research ; Nursing Staff, Hospital/education ; Staff Development/methods ; Surveys and Questionnaires
Language	English
Publishing date	2018-07-05
Publishing country	United States
Document type	Journal Article
ISSN	2169-981X
ISSN (online)	2169-981X
DOI	10.1097/NND.0000000000000462
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Creating resistance to avian influenza infection through genome editing of the ANP32 gene family.

Idoko-Akoh, Alewo / Goldhill, Daniel H / Sheppard, Carol M / Bialy, Dagmara / Quantrill, Jessica L / Sukhova, Ksenia / Brown, Jonathan C / Richardson, Samuel / Campbell, Ciara / Taylor, Lorna / Sherman, Adrian / Nazki, Salik / Long, Jason S / Skinner, Michael A / Shelton, Holly / Sang, Helen M / Barclay, Wendy S / McGrew, Mike J

Nature communications

2023 Volume 14, Issue 1, Page(s) 6136

Abstract: Chickens genetically resistant to avian influenza could prevent future outbreaks. In chickens, influenza A virus (IAV) relies on host protein ANP32A. Here we use CRISPR/Cas9 to generate homozygous gene edited (GE) chickens containing two ANP32A amino ... ...

Abstract	Chickens genetically resistant to avian influenza could prevent future outbreaks. In chickens, influenza A virus (IAV) relies on host protein ANP32A. Here we use CRISPR/Cas9 to generate homozygous gene edited (GE) chickens containing two ANP32A amino acid substitutions that prevent viral polymerase interaction. After IAV challenge, 9/10 edited chickens remain uninfected. Challenge with a higher dose, however, led to breakthrough infections. Breakthrough IAV virus contained IAV polymerase gene mutations that conferred adaptation to the edited chicken ANP32A. Unexpectedly, this virus also replicated in chicken embryos edited to remove the entire ANP32A gene and instead co-opted alternative ANP32 protein family members, chicken ANP32B and ANP32E. Additional genome editing for removal of ANP32B and ANP32E eliminated all viral growth in chicken cells. Our data illustrate a first proof of concept step to generate IAV-resistant chickens and show that multiple genetic modifications will be required to curtail viral escape.
MeSH term(s)	Chick Embryo ; Animals ; Influenza in Birds/genetics ; Gene Editing ; RNA-Dependent RNA Polymerase/genetics ; RNA-Dependent RNA Polymerase/metabolism ; Chickens/genetics ; Influenza A virus/genetics ; Influenza A virus/metabolism
Chemical Substances	RNA-Dependent RNA Polymerase (EC 2.7.7.48)
Language	English
Publishing date	2023-10-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-023-41476-3
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Avian Primordial Germ Cells Are Bipotent for Male or Female Gametogenesis.

Ballantyne, Maeve / Taylor, Lorna / Hu, Tuanjun / Meunier, Dominique / Nandi, Sunil / Sherman, Adrian / Flack, Brenda / Henshall, John M / Hawken, Rachel J / McGrew, Mike J

Frontiers in cell and developmental biology

2021 Volume 9, Page(s) 726827

Abstract: In birds, males are the homogametic sex (ZZ) and females are the heterogametic sex (ZW). Here, we investigate the role of chromosomal sex and germ cell competition on avian germ cell differentiation. We recently developed genetically sterile layer ... ...

Abstract	In birds, males are the homogametic sex (ZZ) and females are the heterogametic sex (ZW). Here, we investigate the role of chromosomal sex and germ cell competition on avian germ cell differentiation. We recently developed genetically sterile layer cockerels and hens for use as surrogate hosts for primordial germ cell (PGC) transplantation. Using
Language	English
Publishing date	2021-09-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2021.726827
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Article ; Online: A low-tech, cost-effective and efficient method for safeguarding genetic diversity by direct cryopreservation of poultry embryonic reproductive cells

Tuanjun Hu / Taylor, Lorna / Sherman, Adrian / Tiambo, Christian K. / Kemp, Stephen J. / Whitelaw, Bruce / Hawken, Rachel J. / Djikeng, Appolinaire / McGrew, Michael J.

eLife

2022

Abstract	Chickens are an important resource for smallholder farmers who raise locally adapted, genetically distinct breeds for eggs and meat. The development of efficient reproductive technologies to conserve and regenerate chicken breeds safeguards existing biodiversity and secures poultry genetic resources for climate resilience, biosecurity, and future food production. The majority of the over 1600 breeds of chicken are raised in low and lower to middle income countries under resource-limited, small-scale production systems, which necessitates a low-tech, cost-effective means of conserving diversity is needed. Here, we validate a simple biobanking technique using cryopreserved embryonic chicken gonads. The gonads are quickly isolated, visually sexed, pooled by sex, and cryopreserved. Subsequently, the stored material is thawed and dissociated before injection into sterile host chicken embryos. By using pooled GFP and RFP-labelled donor gonadal cells and Sire Dam Surrogate mating, we demonstrate that chicks deriving entirely from male and female donor germ cells are hatched. This technology will enable ongoing efforts to conserve chicken genetic diversity for both commercial and smallholder farmers, and to preserve existing genetic resources at poultry research facilities.
Keywords	poultry ; animal breeding ; genetics ; chickens ; cryopreservation
Language	English
Publishing date	2022-01-27T17:38:41Z
Publisher	eLife Sciences Publications, Ltd
Publishing country	fr
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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