A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
A robust knock-in approach using a minimal promoter and a minicircle
Dev Biol. 2023 Oct 13:S0012-1606(23)00170-7. doi: 10.1016/j.ydbio.2023.10.002. Online ahead of print.ABSTRACTKnock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated ...
Source: Developmental Biology - October 15, 2023 Category: Biology Authors: Margaret Keating Ryan Hagle Daniel Osorio-Mendez Anjelica Rodriguez-Parks Sarah I Almutawa Junsu Kang Source Type: research
Exploring the molecular mechanism of coloration differences in two Meconopsis wilsonii subspecies: australis and orientalis
In this study, we collected petals from Australis and Orientalis at different developmental stages to compare the coloration differences between the two species and detect the molecular mechanisms of blue color in Orientalis. We identified that cyanidin was the main anthocyanin in the flowers of both species, and the blue color in Orientalis primarily arises from anthocyanins (Cyanidin-3-O-sambubioside). RNA sequencing analysis was performed to detect the gene expression in the anthocyanin biosynthesis pathway, and the results suggested that gene regulation for anthocyanin biosynthesis may not be the direct reason for blue...
Source: Developmental Biology - October 14, 2023 Category: Biology Authors: Zhi Ou Jun Luo Yan Qu Source Type: research
Exploring the molecular mechanism of coloration differences in two Meconopsis wilsonii subspecies: australis and orientalis
In this study, we collected petals from Australis and Orientalis at different developmental stages to compare the coloration differences between the two species and detect the molecular mechanisms of blue color in Orientalis. We identified that cyanidin was the main anthocyanin in the flowers of both species, and the blue color in Orientalis primarily arises from anthocyanins (Cyanidin-3-O-sambubioside). RNA sequencing analysis was performed to detect the gene expression in the anthocyanin biosynthesis pathway, and the results suggested that gene regulation for anthocyanin biosynthesis may not be the direct reason for blue...
Source: Developmental Biology - October 14, 2023 Category: Biology Authors: Zhi Ou Jun Luo Yan Qu Source Type: research
Loss of Baz1b in mice causes perinatal lethality, growth failure, and variable multi-system outcomes
Dev Biol. 2023 Oct 10:S0012-1606(23)00161-6. doi: 10.1016/j.ydbio.2023.09.007. Online ahead of print.ABSTRACTBAZ1B is one of 25-27 coding genes deleted in canonical Williams syndrome, a multi-system disorder causing slow growth, vascular stenosis, and gastrointestinal complaints, including constipation. BAZ1B is involved in (among other processes) chromatin organization, DNA damage repair, and mitosis, suggesting reduced BAZ1B may contribute to Williams syndrome symptoms. In mice, loss of Baz1b causes early neonatal death. 89.6% of Baz1b-/- mice die within 24 h of birth without vascular anomalies or congenital heart diseas...
Source: Developmental Biology - October 12, 2023 Category: Biology Authors: Christopher Pai Basil A McIntosh Russell H Knutsen Mark D Levin Kit Man Tsang Beth A Kozel Robert O Heuckeroth Source Type: research
Loss of Baz1b in mice causes perinatal lethality, growth failure, and variable multi-system outcomes
Dev Biol. 2023 Oct 10:S0012-1606(23)00161-6. doi: 10.1016/j.ydbio.2023.09.007. Online ahead of print.ABSTRACTBAZ1B is one of 25-27 coding genes deleted in canonical Williams syndrome, a multi-system disorder causing slow growth, vascular stenosis, and gastrointestinal complaints, including constipation. BAZ1B is involved in (among other processes) chromatin organization, DNA damage repair, and mitosis, suggesting reduced BAZ1B may contribute to Williams syndrome symptoms. In mice, loss of Baz1b causes early neonatal death. 89.6% of Baz1b-/- mice die within 24 h of birth without vascular anomalies or congenital heart diseas...
Source: Developmental Biology - October 12, 2023 Category: Biology Authors: Christopher Pai Basil A McIntosh Russell H Knutsen Mark D Levin Kit Man Tsang Beth A Kozel Robert O Heuckeroth Source Type: research
In vivo tracing of the Cytokeratin 14 lineages using self-cleaving guide RNAs and CRISPR/Cas9
Dev Biol. 2023 Oct 7:S0012-1606(23)00165-3. doi: 10.1016/j.ydbio.2023.09.011. Online ahead of print.ABSTRACTThe current gold-standard for genetic lineage tracing in transgenic mice is based on cell-type specific expression of Cre recombinase. As an alternative, we developed a cell-type specific CRISPR/spCas9 system for lineage tracing. This method relies on RNA polymerase II promoter driven self-cleaving guide RNAs (scgRNA) to achieve tissue-specificity. To demonstrate proof-of-principle for this approach a transgenic mouse was generated harbouring a knock-in of a scgRNA into the Cytokeratin 14 (Krt14) locus. Krt14 express...
Source: Developmental Biology - October 9, 2023 Category: Biology Authors: Amita Tiyaboonchai Leslie Wakefield Anne Vonada Catherine L May Craig Dorrell David Enicks Anusha Sairavi Klaus H Kaestner Markus Grompe Source Type: research
In vivo tracing of the Cytokeratin 14 lineages using self-cleaving guide RNAs and CRISPR/Cas9
Dev Biol. 2023 Oct 7:S0012-1606(23)00165-3. doi: 10.1016/j.ydbio.2023.09.011. Online ahead of print.ABSTRACTThe current gold-standard for genetic lineage tracing in transgenic mice is based on cell-type specific expression of Cre recombinase. As an alternative, we developed a cell-type specific CRISPR/spCas9 system for lineage tracing. This method relies on RNA polymerase II promoter driven self-cleaving guide RNAs (scgRNA) to achieve tissue-specificity. To demonstrate proof-of-principle for this approach a transgenic mouse was generated harbouring a knock-in of a scgRNA into the Cytokeratin 14 (Krt14) locus. Krt14 express...
Source: Developmental Biology - October 9, 2023 Category: Biology Authors: Amita Tiyaboonchai Leslie Wakefield Anne Vonada Catherine L May Craig Dorrell David Enicks Anusha Sairavi Klaus H Kaestner Markus Grompe Source Type: research
In vivo tracing of the Cytokeratin 14 lineages using self-cleaving guide RNAs and CRISPR/Cas9
Dev Biol. 2023 Oct 7:S0012-1606(23)00165-3. doi: 10.1016/j.ydbio.2023.09.011. Online ahead of print.ABSTRACTThe current gold-standard for genetic lineage tracing in transgenic mice is based on cell-type specific expression of Cre recombinase. As an alternative, we developed a cell-type specific CRISPR/spCas9 system for lineage tracing. This method relies on RNA polymerase II promoter driven self-cleaving guide RNAs (scgRNA) to achieve tissue-specificity. To demonstrate proof-of-principle for this approach a transgenic mouse was generated harbouring a knock-in of a scgRNA into the Cytokeratin 14 (Krt14) locus. Krt14 express...
Source: Developmental Biology - October 9, 2023 Category: Biology Authors: Amita Tiyaboonchai Leslie Wakefield Anne Vonada Catherine L May Craig Dorrell David Enicks Anusha Sairavi Klaus H Kaestner Markus Grompe Source Type: research
SPIN-4/spinster supports sperm activation in C. elegans via sphingosine-1-phosphate transport
Dev Biol. 2023 Oct 5:S0012-1606(23)00167-7. doi: 10.1016/j.ydbio.2023.09.013. Online ahead of print.ABSTRACTSpermiogenesis, a sperm-activation step, is crucial for the transformation of immotile spermatids into motile sperm. Though membrane transport of ions and molecules across the sperm plasma membrane has been implicated in this process, the full repertoire of transporters involved, and their respective substrates, is unclear. Here, we report that the major facilitator superfamily transporter SPIN-4/Spinster governs efficient spermiogenesis and fertility in the hermaphrodite nematode Caenorhabditis elegans. Unlike other...
Source: Developmental Biology - October 7, 2023 Category: Biology Authors: Yash Flora K Adam Bohnert Source Type: research
The BAF chromatin complex component SMARCC1 does not mediate GLI transcriptional repression of Hedgehog target genes in limb buds
Dev Biol. 2023 Oct 5:S0012-1606(23)00169-0. doi: 10.1016/j.ydbio.2023.10.001. Online ahead of print.ABSTRACTTranscriptional responses to the Hedgehog (HH) signaling pathway are primarily modulated by GLI repression in the mouse limb. Previous studies suggested a role for the BAF chromatin remodeling complex in mediating GLI repression. Consistent with this possibility, the core BAF complex protein SMARCC1 is present at most active limb enhancers including the majority of GLI enhancers. However, in contrast to GLI repression which reduces chromatin accessibility, SMARCC1 maintains chromatin accessibility at most enhancers, ...
Source: Developmental Biology - October 7, 2023 Category: Biology Authors: Janani Ramachandran Wanlu Chen Rachel K Lex Kathryn E Windsor Hyunji Lee Tingchang Wang Weiqiang Zhou Hongkai Ji Steven A Vokes Source Type: research
