Paul Verma | Monash University (original) (raw)

Papers by Paul Verma

bioRxiv (Cold Spring Harbor Laboratory), Jun 24, 2023

Journal of Equine Veterinary Science, Jul 1, 2018

Elsevier eBooks, 2021

Abstract Cattle constitute one of the most commercially important livestock species. They are a s... more Abstract Cattle constitute one of the most commercially important livestock species. They are a significant source of nutrition as well has had great economic importance. Through thousands of years of selective breeding humans have shaped cattle into these multipurpose species that are adapted to various environments around the globe. In the past decade, the sequencing of the cattle genome has paved the way for genetic enhancement of existing breeds to increase productivity and sustainability. More recently, developments in genome editing technologies and pluripotent stem cell culture can now be combined to achieve highly commercial goals for the livestock industry. In this chapter, we discuss the basics of these cutting-edge technologies and their applications in cattle. We focus on bovine iPSCs (biPSCs) as they can be generated in theory from any individual long after their genetic value has been proven, and their phenotypic characteristics validated and regardless of their fecundity status. Furthermore, we discuss the various genome editors and how these novel tools can be used for the genetic improvement of cattle.

bioRxiv (Cold Spring Harbor Laboratory), Feb 19, 2023

Humana Press eBooks, 2006

To produce autologous ESCs for a bovine model of cell therapy, we activated oocytes by calcium io... more To produce autologous ESCs for a bovine model of cell therapy, we activated oocytes by calcium ionophore (CI) and 6 dimethylaminopurine (6 DMAP) and isolated ESCs from the resulting parthenotes. Parthenote ESC lines (pbESC) would also provide a valuable tool for epigenetic studies on ESCs. Five pbESC like-cell lines were expanded for 12 passages over 120 days and differentiated to form embryoid bodies by suspension culture. The pbESC lines demonstrated typical ESC morphology and expressed ESC markers including alkaline phosphate and stage-specific embryonic antigen, SSEA1 and SSAE4 asssessed by histochemical and immuno-fluorescence staining, respectively. In addition, gene expression of Oct4, Rex1, SSEA1 and ALP was confirmed using RT–PCR. These cells had a normal karyotype. The cells formed EBs and showed expression of the markers of three embryonic germ layers. In summary, we show that ESCs can be derived from bovine parthenogenetic blastocysts and that these cells express pluripotent markers and have ability to form EBs and differentiate into cells indicative of the three embryonic germ layers. Additional work will focus on imprinted gene expression and will provide further evidence of the parthenogenetic origin of the pbESC lines.

Zenodo (CERN European Organization for Nuclear Research), Jun 19, 2023

Stem Cells International, 2018

Mesenchymal stromal cell-like populations have been derived from mouse-induced pluripotent stem c... more Mesenchymal stromal cell-like populations have been derived from mouse-induced pluripotent stem cells (miPSC-MSC) with the capability for tissue regeneration. In this study, murine iPSC underwent differentiation towards an MSC-like immunophenotype. Stable miPSC-MSC cultures expressed the MSC-associated markers, CD73, CD105, and Sca-1, but lacked expression of the pluripotency marker, SSEA1, and hematopoietic markers, CD34 and CD45. Functionally, miPSC-MSC exhibited the potential for trilineage differentiation into osteoblasts, adipocytes, and chondrocytes and the capacity to suppress the proliferation of mitogen-activated splenocytes. The efficacy of miPSC-MSC was assessed in an acute inflammation model following systemic or local delivery into mice with subcutaneous implants containing heat-inactivated P. gingivalis. Histological analysis revealed less inflammatory cellular infiltrate within the sponges in mice treated with miPSC-MSC cells delivered locally rather than systemically...

Reproduction, Fertility and Development, 2005

The efficiency of obtaining live calves from somatic cell nuclear transfer remains quite low. One... more The efficiency of obtaining live calves from somatic cell nuclear transfer remains quite low. One factor implicated in this failure is inadequate chromatin remodelling of the donor nucleus. Five Polycomb group (PcG) genes were investigated as potential remodelling factors in bovine oocytes and preimplantation embryos. These genes (Cbx6, Eed, Edr1, Yy1, and Zfp144) are involved in transcriptional activation and cell cycle regulation. We hypothesize that inadequate expression may cause the developmental abnormalities seen following cloning. This study is aimed at characterizing normal expression, prior to comparative studies with cloned embryos. Three single abattoir-derived in vitro-matured (IVM) oocytes or in vitro-produced (IVP) embryos from each of the following stages: 2-cell, 4-cell, 8-cell, 16–32 cell, morula, Day 7 blastocyst, and Day 8 hatched blastocyst, were studied. Messenger RNA was isolated from individual samples with Dynabeads (Dynal, Inc., Lake Success, NY, USA) and t...

<p>(A) Illustration of full-length viral glycoprotein-1 (GP1) of Lymphocytic Choriomeningit... more <p>(A) Illustration of full-length viral glycoprotein-1 (GP1) of Lymphocytic Choriomeningitis Virus (LCMV) to which Transduction Domains (TD)-1, TD-2, TD-1.1, TD-2.1 & TD-2.2 were devised. Blue bars represent domains within the amino acid sequence of GP1 that are conserved with other a-dystroglycan binding proteins (Laminin2 and Agrin; see B for protein sequence alignments). TD’s are devised from these conserved regions. (B) Protein sequence alignments of GP1 and Laminin-2 or Agrin. Amino acid domains of Laminin-2 or Agrin are denoted and numbered to each side. TD1 and TD2 are identical to GP1 domains. TD1.1, TD2.1 & TD2.2 are composite sequences of GP1/Laminin-2 and Agrin; residues were included if any of the Laminin-2 or Agrin conserved sequences had identical amino acids to the reference GP1 sequence (shown in black). Where identical residues could not be found, amino acids represented more than once among the Laminin-2 and Agrin sequences were included (shown in red). Where no similarity was observed between reference GP1 and Laminin-2/Agrin sequences, cationic residues (R, K) were included for their transmembrane transduction abilities (in normal type; reviewed <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045501#pone.0045501-Sawant1" target="_blank">[35]</a>). (C) Diagrammatic outline of the transduction domain constructs. TD1.1, TD-2.1 & TD-2.2 are fused to a histidine leader sequence. TD1 was fused to EGFP (interspaced by a flexible -SGGGS- linker sequence) and a histidine leader sequence. Right are predicted molecular weights (Dalton) and isoelectric points (pI) of pET-TD proteins (MW & pI both ascertained with ExPasy software; <a href="http://ca.expasy.org/tools/pi_tool.html" target="_blank">http://ca.expasy.org/tools/pi_tool.html</a>).</p

Methods in molecular biology, 2022

Reproduction, Fertility and Development, 2008

Frontiers in Cell and Developmental Biology

Somatic cell nuclear transfer (SCNT) is a key technology with broad applications that range from ... more Somatic cell nuclear transfer (SCNT) is a key technology with broad applications that range from production of cloned farm animals to derivation of patient-matched stem cells or production of humanized animal organs for xenotransplantation. However, effects of aberrant epigenetic reprogramming on gene expression compromise cell and organ phenotype, resulting in low success rate of SCNT. Standard SCNT procedures include enucleation of recipient oocytes before the nuclear donor cell is introduced. Enucleation removes not only the spindle apparatus and chromosomes of the oocyte but also the perinuclear, mitochondria rich, ooplasm. Here, we use a Bos taurus SCNT model with in vitro fertilized (IVF) and in vivo conceived controls to demonstrate a ∼50% reduction in mitochondrial DNA (mtDNA) in the liver and skeletal muscle, but not the brain, of SCNT fetuses at day 80 of gestation. In the muscle, we also observed significantly reduced transcript abundances of mtDNA-encoded subunits of the...

Copyright © 2012 Khodadad Khodadadi et al. This is an open access article distributed under the C... more Copyright © 2012 Khodadad Khodadadi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Despite tremendous efforts on isolation of pluripotent equine embryonic stem (ES) cells, to date there are few reports about successful isolation of ESCs and no report of in vivo differentiation of this important companion species. We report the induction of pluripotency in adult equine fibroblasts via retroviral transduction with three transcription factors using OCT4, SOX2, and KLF4 in the absence of c-MYC. The cell lines were maintained beyond 27 passages (more than 11 months) and characterized. The equine iPS (EiPS) cells stained positive for alkaline phosphatase by histochemical staining and expressed OCT4, NANOG, SSEA1, and SSEA4. Gene expression analysis of the cells showed the expression of OCT4, SOX2 NANOG, andSTAT3. The...

Methods in Molecular Biology, 2015

Fertilization triggers a cascade of cellular and molecular events restoring the totipotent state ... more Fertilization triggers a cascade of cellular and molecular events restoring the totipotent state and the potential for all cell types. However, the program quickly directs differentiation and cellular commitment. Under the genetic and epigenetic control of this process, Waddington likened this to a three-dimensional landscape where cells could not ascend the slope or traverse once canalized thus leading to cell fate decisions and the progressive restriction of cellular potency. But this is not the only possible outcome at least experimentally. Somatic cell nuclear transfer and overexpression of key transcription factors to generate induced pluripotent cells have challenged this notion. The return to pluripotency and the reinstatement of plasticity and heterogeneity once thought to be the exclusive remit of the developing embryo can now be replicated in vitro. The following chapter introduces some of these ideas and suggests that the fundamental principles learned may constitute the fi rst step toward the opportunity for specifi c tissue renewal and replacement in healthy aging and the treatment of chronic diseases—the age of regenerative medicine.