Efficient Generation of SOCS2 Knock-Out Sheep by Electroporation of CRISPR-Cas9 Ribonucleoprotein Complex with Dual-sgRNAs Ahmed K. Mahdi, Devon S. Fitzpatrick, Darren E. Hagen, Bret R. McNabb, Tara Urbano Beach, William M. Muir, Nicholas Werry, Alison L. Van Eenennaam, Juan F. Medrano, Pablo J. Ross Crispr Journal, 2025 In mice, naturally occurring and induced mutations in the suppressor of cytokine signaling-2 ( Socs2 ) gene are associated with a high growth phenotype characterized by rapid post-weaning weight gain and 30–50% heavier mature body weight. In this work, we demonstrate an electroporation-based method of producing SOCS2 knock-out (KO) sheep. Electroporation of dual-guide CRISPR-Cas9 ribonucleoprotein complexes targeting SOCS2 was performed 6 h post-fertilization in sheep zygotes. Fifty-two blastocysts were transferred to 13 estrus-synchronized recipients, yielding five live lambs and one stillborn. These lambs all carried mutations predicted to result in SOCS2 KO. Three carried large deletion alleles which evaded detection in initial PCR screening. Off-target analysis using whole genome sequencing comparing the frequency of mutations in regions within 100 bp of possible sgRNA binding sites (up to 4 bp mismatches) and elsewhere in the genome showed no significant difference when comparing unedited control sheep to edited animals ( p = 0.71). In conclusion, electroporation of zygotes with dual-guide CRISPR-Cas9 RNPs was effective at generating knock-out sheep with no substantial off-target activity.
THE PSEUDOBRANCH DEVELOPMENT IN EASTERN MOSQUITO FISH GAMBUSIA HOLBROOKI, GIRARD 1859 (CYPRINODONTIFORMES, POECILIIDAE) FROM JADIRIYAH LAKE, BAGHDAD, IRAQ Department of Anatomy, Faculty of Medicine, University of Diyala, Diyala, Iraq, Khloud AL-Lehaibe, Entekhab AL-Shuwaili, Department of Biology, College of Education for Pure Science (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq, Ahmed Mahdi, Department of Biology, College of Education for Pure Science (Ibn Al-Haitham), University of Baghdad, Baghdad, Iraq Bulletin of the Iraq Natural History Museum, 2025 The aim of the current investigation is to study the embryogenesis of the pseudobranchia (PB) in the Eastern mosquito fish Gambusia holbrooki, Girard, 1859 (Cyprinodontiformes, Poeciliidae). Histological sections from 2-4 mm long embryos revealed the presumable PB as a cell mass surrounded by a row of squamous cells in the operculum, as well as the formation of cartilage and blood vessels, and the beginning of the formation of its lamellae. In 5-7.75 mm long embryos, a prominent rise appeared in the mass of cells and PB lamellae, along with an increase in melanin deposition around the blood vessels. Moreover, a row of squamous cells encircled the rise from the front and back. At this stage, the parallel arrangement of the cartilage-attached lamellae correlated with increased melanin concentration around the blood vessels. In newborns, the differentiated cells increased similar to that appearing in the adult’s stages.
Single-Step Genome Editing of Small Ruminant Embryos by Electroporation Ahmed K. Mahdi, Juan F. Medrano, Pablo J. Ross International Journal of Molecular Sciences, 2022 We investigated the possibility of single-step genome editing in small ruminants by CRISPR-Cas9 zygote electroporation. We targeted SOCS2 and PDX1 in sheep embryos and OTX2 in goat embryos, utilizing a dual sgRNA approach. Gene editing efficiency was compared between microinjection and three different electroporation settings performed at four different times of embryo development. Electroporation of sheep zygotes 6 h after fertilization with settings that included short high-voltage (poring) and long low-voltage (transfer) pulses was efficient at producing SOCS2 knock-out blastocysts. The mutation rate after CRISPR/Cas9 electroporation was 95.6% ± 8%, including 95.4% ± 9% biallelic mutations; which compared favorably to 82.3% ± 8% and 25% ± 10%, respectively, when using microinjection. We also successfully disrupted the PDX1 gene in sheep and the OTX2 gene in goat embryos. The biallelic mutation rate was 81 ± 5% for PDX1 and 85% ± 6% for OTX2. In conclusion, using single-step CRISPR-Cas9 zygote electroporation, we successfully introduced biallelic deletions in the genome of small ruminant embryos.
WIP1 Inhibition by GSK2830371 Potentiates HDM201 Through Enhanced p53 Phosphorylation and Activation in Liver Adenocarcinoma Cells.(vol 13, 3876, 2021) CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... CANCERS 18 (3) , 2026 2026
Correction: Wu et al. WIP1 Inhibition by GSK2830371 Potentiates HDM201 Through Enhanced p53 Phosphorylation and Activation in Liver Adenocarcinoma Cells. Cancers 2021, 13, 3876 CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... Cancers 18 (3), 530 , 2026 2026
Key features of the environment promoting liver cancer in the absence of cirrhosis MYW Zaki, AK Mahdi, GL Patman, A Whitehead, JP Mauricio, MV McCain, ... Scientific reports 11 (1), 16727 , 2021 2021 Citations: 18
WIP1 inhibition by GSK2830371 potentiates HDM201 through enhanced p53 phosphorylation and activation in liver adenocarcinoma cells CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... Cancers 13 (15), 3876 , 2021 2021 Citations: 14
OP-01 Genome wide RNA expression analysis identifies CD44 positive macrophages as promoters of hepatocyte proliferation and the development of NAFLD-HCC HLR Marco Youssef William Zaki, Misti V McCain, Ahmed K Mahdi, Joao Mauricio ... EASL HCC summit 2019 , 2019 2019
What drives development of HCC in non-cirrhotic NAFLD? A Mahdi, M Zaki, J Lunec, Q Anstee, D Tiniakos, R Shukla, F Oakley, ... Journal of Hepatology 68, S49 , 2018 2018
Targeting negative regulation of p53 by MDM2 and WIP1 as a therapeutic strategy in cutaneous melanoma CE Wu, A Esfandiari, YH Ho, N Wang, AK Mahdi, E Aptullahoglu, P Lovat, ... British journal of cancer 118 (4), 495-508 , 2018 2018 Citations: 68
1. LIQUID BIOPSY-A NEW PROSPECT AK Mahdi Iraqi Journal of Medical Sciences 16 (4) , 2018 2018
Molecular pathology: the roles of P53 in the oxidative stress and DNA damage responses in chronic liver disease and hepatocellular carcinoma AK Mahdi Newcastle University , 2018 2018 Citations: 2
Lipogranulomas, neutrophils and the progression of NAFLD to NASH and HCC JC Mauricio, AK Mahdi, J Lunec, G Patman, A Whitehead, D Tiniakos, ... HEPATOLOGY 66, 987A-988A , 2017 2017
MDM2 inhibition for non-genotoxic activation of TP53-a novel strategy for the treatment of hepatocellular carcinoma (HCC) AK Mahdi, HD Thomas, S Wedge, R Shukla, H Reeves, J Lunec HEPATOLOGY 66, 369A-369A , 2017 2017
Inhibition of WIP1/PPM1D phosphatase by GSK2830371 potentiates the growth inhibitory and cytotoxic activity of MDM2 antagonists (nutlin-3, RG7388 and HDM201) in cutaneous … CE Wu, A Esfandiari, YH Ho, C Shepherd, AK Mahdi, E Aptullahoglu, ... Cancer Research 77 (13_Supplement), 2151-2151 , 2017 2017 Citations: 1
Abstract P3-07-21: Cytotoxic potential of the RG7388 MDM2-p53 binding antagonist and the GSK2830371 WIP1 inhibitor on MX-1 and MCF-7 human breast cancer cells V Manoharan, J Lunec, A Esfandiari, A Mahdi, CE Wu, M Zanjirband, ... Cancer Research 77 (4_Supplement), P3-07-21-P3-07-21 , 2017 2017 Citations: 2
Cytotoxic potential of the RG7388 MDM2-p53 binding antagonist and the GSK2830371 WIP1 inhibitor on MX-1 and MCF-7 human breast cancer cells V Manoharan, J Lunec, A Esfandiari, A Mahdi, CE Wu, M Zanjirband, ... CANCER RESEARCH 77 , 2017 2017
MDM2 antagonists− a therapeutic approach for patients with hepatocellular carcinoma? A Mahdi, J Lunec, H Reeves European Journal of Cancer 1 (61), S44 , 2016 2016
Bcl-2 Expression in CagA Strain H. Pylori Gastritis (Immunohistochemical and Insitu Hybridization Study) HH Ali Iraqi Postgraduate Medical Journal 11 (1), 12 , 2012 2012
Iraqi JMS TW Ali, AK Mahdi, HH Ali, HA Hassan IRAQI JOURNAL OF MEDICAL SCIENCES, 160 , 2000 2000
MOST CITED SCHOLAR PUBLICATIONS
Targeting negative regulation of p53 by MDM2 and WIP1 as a therapeutic strategy in cutaneous melanoma CE Wu, A Esfandiari, YH Ho, N Wang, AK Mahdi, E Aptullahoglu, P Lovat, ... British journal of cancer 118 (4), 495-508 , 2018 2018 Citations: 68
Key features of the environment promoting liver cancer in the absence of cirrhosis MYW Zaki, AK Mahdi, GL Patman, A Whitehead, JP Mauricio, MV McCain, ... Scientific reports 11 (1), 16727 , 2021 2021 Citations: 18
WIP1 inhibition by GSK2830371 potentiates HDM201 through enhanced p53 phosphorylation and activation in liver adenocarcinoma cells CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... Cancers 13 (15), 3876 , 2021 2021 Citations: 14
Molecular pathology: the roles of P53 in the oxidative stress and DNA damage responses in chronic liver disease and hepatocellular carcinoma AK Mahdi Newcastle University , 2018 2018 Citations: 2
Abstract P3-07-21: Cytotoxic potential of the RG7388 MDM2-p53 binding antagonist and the GSK2830371 WIP1 inhibitor on MX-1 and MCF-7 human breast cancer cells V Manoharan, J Lunec, A Esfandiari, A Mahdi, CE Wu, M Zanjirband, ... Cancer Research 77 (4_Supplement), P3-07-21-P3-07-21 , 2017 2017 Citations: 2
Inhibition of WIP1/PPM1D phosphatase by GSK2830371 potentiates the growth inhibitory and cytotoxic activity of MDM2 antagonists (nutlin-3, RG7388 and HDM201) in cutaneous … CE Wu, A Esfandiari, YH Ho, C Shepherd, AK Mahdi, E Aptullahoglu, ... Cancer Research 77 (13_Supplement), 2151-2151 , 2017 2017 Citations: 1
WIP1 Inhibition by GSK2830371 Potentiates HDM201 Through Enhanced p53 Phosphorylation and Activation in Liver Adenocarcinoma Cells.(vol 13, 3876, 2021) CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... CANCERS 18 (3) , 2026 2026
Correction: Wu et al. WIP1 Inhibition by GSK2830371 Potentiates HDM201 Through Enhanced p53 Phosphorylation and Activation in Liver Adenocarcinoma Cells. Cancers 2021, 13, 3876 CE Wu, AK Mahdi, CY Huang, CP Chen, YR Pan, JWC Chang, JS Chen, ... Cancers 18 (3), 530 , 2026 2026
OP-01 Genome wide RNA expression analysis identifies CD44 positive macrophages as promoters of hepatocyte proliferation and the development of NAFLD-HCC HLR Marco Youssef William Zaki, Misti V McCain, Ahmed K Mahdi, Joao Mauricio ... EASL HCC summit 2019 , 2019 2019
What drives development of HCC in non-cirrhotic NAFLD? A Mahdi, M Zaki, J Lunec, Q Anstee, D Tiniakos, R Shukla, F Oakley, ... Journal of Hepatology 68, S49 , 2018 2018
1. LIQUID BIOPSY-A NEW PROSPECT AK Mahdi Iraqi Journal of Medical Sciences 16 (4) , 2018 2018
Lipogranulomas, neutrophils and the progression of NAFLD to NASH and HCC JC Mauricio, AK Mahdi, J Lunec, G Patman, A Whitehead, D Tiniakos, ... HEPATOLOGY 66, 987A-988A , 2017 2017
MDM2 inhibition for non-genotoxic activation of TP53-a novel strategy for the treatment of hepatocellular carcinoma (HCC) AK Mahdi, HD Thomas, S Wedge, R Shukla, H Reeves, J Lunec HEPATOLOGY 66, 369A-369A , 2017 2017
Cytotoxic potential of the RG7388 MDM2-p53 binding antagonist and the GSK2830371 WIP1 inhibitor on MX-1 and MCF-7 human breast cancer cells V Manoharan, J Lunec, A Esfandiari, A Mahdi, CE Wu, M Zanjirband, ... CANCER RESEARCH 77 , 2017 2017
MDM2 antagonists− a therapeutic approach for patients with hepatocellular carcinoma? A Mahdi, J Lunec, H Reeves European Journal of Cancer 1 (61), S44 , 2016 2016
Bcl-2 Expression in CagA Strain H. Pylori Gastritis (Immunohistochemical and Insitu Hybridization Study) HH Ali Iraqi Postgraduate Medical Journal 11 (1), 12 , 2012 2012
Iraqi JMS TW Ali, AK Mahdi, HH Ali, HA Hassan IRAQI JOURNAL OF MEDICAL SCIENCES, 160 , 2000 2000
