The Experts below are selected from a list of 213 Experts worldwide ranked by ideXlab platform
Rajesh V Thakker - One of the best experts on this subject based on the ideXlab platform.
-
Animal models of pituitary neoplasia.
Molecular and Cellular Endocrinology, 2015Co-Authors: Kate E Lines, Mark Stevenson, Rajesh V ThakkerAbstract:Pituitary neoplasias can occur as part of a complex inherited disorder, or more commonly as sporadic (non-familial) disease. Studies of the molecular and Genetic mechanisms causing such pituitary tumours have identified dysregulation of >35 Genes, with many revealed by studies in mice, rats and zebrafish. Strategies used to Generate these animal models have included Gene knockout, Gene Knockin and transgenic over-expression, as well as chemical mutaGenesis and drug induction. These animal models provide an important resource for investigation of tissue-specific tumourigenic mechanisms, and evaluations of novel therapies, illustrated by studies into multiple endocrine neoplasia type 1 (MEN1), a hereditary syndrome in which ∼30% of patients develop pituitary adenomas. This review describes animal models of pituitary neoplasia that have been Generated, together with some recent advances in Gene editing technologies, and an illustration of the use of the Men1 mouse as a pre clinical model for evaluating novel therapies.
-
Animal models of pituitary neoplasia.
Molecular and Cellular Endocrinology, 2015Co-Authors: Kate E Lines, Mark Stevenson, Rajesh V ThakkerAbstract:Pituitary neoplasias can occur as part of a complex inherited disorder, or more commonly as sporadic (non-familial) disease. Studies of the molecular and Genetic mechanisms causing such pituitary tumours have identified dysregulation of >35 Genes, with many revealed by studies in mice, rats and zebrafish. Strategies used to Generate these animal models have included Gene knockout, Gene Knockin and transgenic over-expression, as well as chemical mutaGenesis and drug induction. These animal models provide an important resource for investigation of tissue-specific tumourigenic mechanisms, and evaluations of novel therapies, illustrated by studies into multiple endocrine neoplasia type 1 (MEN1), a hereditary syndrome in which ∼30% of patients develop pituitary adenomas. This review describes animal models of pituitary neoplasia that have been Generated, together with some recent advances in Gene editing technologies, and an illustration of the use of the Men1 mouse as a pre clinical model for evaluating novel therapies.
Bhanu Prakash V L Telugu - One of the best experts on this subject based on the ideXlab platform.
-
targeted Gene Knockin in porcine somatic cells using crispr cas ribonucleoproteins
International Journal of Molecular Sciences, 2016Co-Authors: Kieun Park, Chihun Park, Anne M Powell, Jessica Martin, David M Donovan, Bhanu Prakash V L TeluguAbstract:The pig is an ideal large animal model for Genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for Generating and propagating Genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in Generating desired Genetic modifications such as targeted Knockin of short stretches of sequences or transGenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted Knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A Gene in porcine somatic cells and Generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these Knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transGene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated Knockin in porcine somatic cells, while simultaneously creating a stable platform for future transGene integration and Generating transgenic animals.
-
Targeted Gene Knockin in Porcine Somatic Cells Using CRISPR/Cas Ribonucleoproteins
International journal of molecular sciences, 2016Co-Authors: Kieun Park, Chihun Park, Anne M Powell, Jessica Martin, David M Donovan, Bhanu Prakash V L TeluguAbstract:The pig is an ideal large animal model for Genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for Generating and propagating Genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in Generating desired Genetic modifications such as targeted Knockin of short stretches of sequences or transGenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted Knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A Gene in porcine somatic cells and Generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these Knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transGene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated Knockin in porcine somatic cells, while simultaneously creating a stable platform for future transGene integration and Generating transgenic animals.
Haruhiko Fujiwara - One of the best experts on this subject based on the ideXlab platform.
-
Targeted Gene Knockin in zebrafish using the 28S rDNA-specific non-LTR-retrotransposon R2Ol
Mobile DNA, 2019Co-Authors: Azusa Kuroki-kami, Narisu Nichuguti, Haruka Yatabe, Sayaka Mizuno, Shoji Kawamura, Haruhiko FujiwaraAbstract:Background Although most of long interspersed elements (LINEs), one class of non-LTR-retrotransposons, are integrated into the host genome randomely, some elements are retrotransposed into the specific sequences of the genomic regions, such as rRNA Gene (rDNA) clusters, telomeric repeats and other repetitive sequenes. Most of the sequence-specific LINEs have been reported mainly among invertebrate species and shown to retrotranspose into the specific sequences in vivo and in vitro systems. Recenlty, 28S rDNA-specific LINE R2 elements are shown to be distributed among widespread vertebrate species, but the sequence-specific retrotransposition of R2 has never been demonstrated in vertebrates. Results Here we cloned a full length unit of R2 from medaka fish Oryzias latipes , named R2Ol, and engineered it to a targeted Gene integration tool in zebrafish. By injecting R2Ol-encoding mRNA into zebrafish embryos, R2Ol retrotransposed precisely into the target site at high efficiency (98%) and was transmitted to the next Generation at high frequency (50%). We also Generated transgenic zebrafish carrying the enhanced green fluorescent protein (EGFP) reporter Gene in 28S rDNA target by the R2Ol retrotransposition system. Conclusions Sequence-specific LINE retrotransposes into the precise sequence using target primed reverse transcription (TPRT), possibly providing an alternative and effective targeted Gene Knockin method in vertebrates.
-
Targeted Gene Knockin in zebrafish using the 28S rDNA-specific non-LTR-retrotransposon R2Ol.
Mobile DNA, 2019Co-Authors: Azusa Kuroki-kami, Narisu Nichuguti, Haruka Yatabe, Sayaka Mizuno, Shoji Kawamura, Haruhiko FujiwaraAbstract:Although most of long interspersed elements (LINEs), one class of non-LTR-retrotransposons, are integrated into the host genome randomely, some elements are retrotransposed into the specific sequences of the genomic regions, such as rRNA Gene (rDNA) clusters, telomeric repeats and other repetitive sequenes. Most of the sequence-specific LINEs have been reported mainly among invertebrate species and shown to retrotranspose into the specific sequences in vivo and in vitro systems. Recenlty, 28S rDNA-specific LINE R2 elements are shown to be distributed among widespread vertebrate species, but the sequence-specific retrotransposition of R2 has never been demonstrated in vertebrates. Here we cloned a full length unit of R2 from medaka fish Oryzias latipes, named R2Ol, and engineered it to a targeted Gene integration tool in zebrafish. By injecting R2Ol-encoding mRNA into zebrafish embryos, R2Ol retrotransposed precisely into the target site at high efficiency (98%) and was transmitted to the next Generation at high frequency (50%). We also Generated transgenic zebrafish carrying the enhanced green fluorescent protein (EGFP) reporter Gene in 28S rDNA target by the R2Ol retrotransposition system. Sequence-specific LINE retrotransposes into the precise sequence using target primed reverse transcription (TPRT), possibly providing an alternative and effective targeted Gene Knockin method in vertebrates.
-
Additional file 1: of Targeted Gene Knockin in zebrafish using the 28S rDNA-specific non-LTR-retrotransposon R2Ol
2019Co-Authors: Azusa Kuroki-kami, Narisu Nichuguti, Haruka Yatabe, Sayaka Mizuno, Shoji Kawamura, Haruhiko FujiwaraAbstract:Figure S1. Comparison of R2 element insertion site. Figure S2. Nine clones of R2Ol obtained from medaka fish genomic DNA. Figure S3. A scheme for transGenesis in zebrafish using R2Ol. Figure S4. F1 progeny analysis of two founder lines, R2f-9 and R2f-10. Figure S5. Experimental approach for analysis of hsp-GAL4:UAS-EGFP expression induced by heat shock. (PDF 836 kb
Kate E Lines - One of the best experts on this subject based on the ideXlab platform.
-
Animal models of pituitary neoplasia.
Molecular and Cellular Endocrinology, 2015Co-Authors: Kate E Lines, Mark Stevenson, Rajesh V ThakkerAbstract:Pituitary neoplasias can occur as part of a complex inherited disorder, or more commonly as sporadic (non-familial) disease. Studies of the molecular and Genetic mechanisms causing such pituitary tumours have identified dysregulation of >35 Genes, with many revealed by studies in mice, rats and zebrafish. Strategies used to Generate these animal models have included Gene knockout, Gene Knockin and transgenic over-expression, as well as chemical mutaGenesis and drug induction. These animal models provide an important resource for investigation of tissue-specific tumourigenic mechanisms, and evaluations of novel therapies, illustrated by studies into multiple endocrine neoplasia type 1 (MEN1), a hereditary syndrome in which ∼30% of patients develop pituitary adenomas. This review describes animal models of pituitary neoplasia that have been Generated, together with some recent advances in Gene editing technologies, and an illustration of the use of the Men1 mouse as a pre clinical model for evaluating novel therapies.
-
Animal models of pituitary neoplasia.
Molecular and Cellular Endocrinology, 2015Co-Authors: Kate E Lines, Mark Stevenson, Rajesh V ThakkerAbstract:Pituitary neoplasias can occur as part of a complex inherited disorder, or more commonly as sporadic (non-familial) disease. Studies of the molecular and Genetic mechanisms causing such pituitary tumours have identified dysregulation of >35 Genes, with many revealed by studies in mice, rats and zebrafish. Strategies used to Generate these animal models have included Gene knockout, Gene Knockin and transgenic over-expression, as well as chemical mutaGenesis and drug induction. These animal models provide an important resource for investigation of tissue-specific tumourigenic mechanisms, and evaluations of novel therapies, illustrated by studies into multiple endocrine neoplasia type 1 (MEN1), a hereditary syndrome in which ∼30% of patients develop pituitary adenomas. This review describes animal models of pituitary neoplasia that have been Generated, together with some recent advances in Gene editing technologies, and an illustration of the use of the Men1 mouse as a pre clinical model for evaluating novel therapies.
Kieun Park - One of the best experts on this subject based on the ideXlab platform.
-
targeted Gene Knockin in porcine somatic cells using crispr cas ribonucleoproteins
International Journal of Molecular Sciences, 2016Co-Authors: Kieun Park, Chihun Park, Anne M Powell, Jessica Martin, David M Donovan, Bhanu Prakash V L TeluguAbstract:The pig is an ideal large animal model for Genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for Generating and propagating Genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in Generating desired Genetic modifications such as targeted Knockin of short stretches of sequences or transGenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted Knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A Gene in porcine somatic cells and Generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these Knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transGene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated Knockin in porcine somatic cells, while simultaneously creating a stable platform for future transGene integration and Generating transgenic animals.
-
Targeted Gene Knockin in Porcine Somatic Cells Using CRISPR/Cas Ribonucleoproteins
International journal of molecular sciences, 2016Co-Authors: Kieun Park, Chihun Park, Anne M Powell, Jessica Martin, David M Donovan, Bhanu Prakash V L TeluguAbstract:The pig is an ideal large animal model for Genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for Generating and propagating Genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in Generating desired Genetic modifications such as targeted Knockin of short stretches of sequences or transGenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted Knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A Gene in porcine somatic cells and Generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these Knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transGene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated Knockin in porcine somatic cells, while simultaneously creating a stable platform for future transGene integration and Generating transgenic animals.