Genome sequencing of Plasmodium malariae identifies continental segregation and mutations associated with reduced pyrimethamine susceptibility by Amy Ibrahim, Franziska Mohring, Emilia Manko, Donelly A. van Schalkwyk, Jody E. Phelan, Debbie Nolder, Steffen Borrmann, Ayola A. Adegnika, Silvia Maria Di Santi, Mohammad Shafiul Alam, Dinesh Mondal, Francois Nosten, Colin J. Sutherland, Robert W. Moon, Taane G. Clark, Susana Campino

“…Additionally, we characterise pmdhfr mutation F57L and demonstrate its involvement in reduced susceptibility to pyrimethamine in an in vitro parasite assay. We validate CRISPR-Cas9 mediated ortholog replacement in P. knowlesi parasites to determine the function of pmdhfr mutations and demonstrate that circulating pmdhfr genotypes are less susceptible to pyrimethamine.…”
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BRCA1 is involved in sustaining rapid antler growth possibly via balancing of the p53/endoplasmic reticulum stress signaling pathway by Qianqian Guo, Zhen Wang, Jiping Li, Chao Ma, Junjun Zheng, Hengxing Ba, Guokun Zhang, Chunyi Li

“…Among these 39 genes, the tumor suppressor gene, BRCA1, was found to be more sensitive to DNA damage in the AnSCs compared to that in the deer somatic cells, and BRCA1 deletion in the AnSCs via CRISPR/Cas9 resulted in significantly higher levels of DNA damage. …”
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Development of ptxD/Phi as a new dominant selection system for genetic manipulation in Cryptococcus neoformans by Muthita Khongthongdam, Tanaporn Phetruen, Sittinan Chanarat

“…Using the transient CRISPR-Cas9 coupled with electroporation system, we integrated the ptxD gene into the C. neoformans genome and assessed the impact of ptxD integration on cell growth and virulence factors. …”
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TaWI12 may be involved in pistillody and leaf cracking in wheat by Yuhuan Guo, Yan Zhang, Yuhao Li, Yichao Wu, Mingli Liao, Zhengsong Peng, Zaijun Yang, Yonghong Zhou

“…Simultaneously, we used the CRISPR/Cas9 system to generate mutant plants via the modification of three homoeologs of TaWI12 in wheat. …”
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Binding of HCN channels to GABAB receptors in dopamine neurons of the VTA limits synaptic inhibition and prevents the development of anxiety by Enrique Pérez-Garci, Kateryna Pysanenko, Giorgio Rizzi, Florian Studer, Daniel Ulrich, Thorsten Fritzius, Simon Früh, Alessandra Porcu, Valérie Besseyrias, Adolf Melichar, Martin Gassmann, Tania Rinaldi Barkat, Rostislav Tureček, Kelly R. Tan, Bernhard Bettler

“…KCTD16−/− mice exhibit increased anxiety-like behavior in response to stress - a behavior replicated by CRISPR/Cas9-mediated KCTD16 ablation in DAVTA neurons or by intra-VTA infusion of an HCN antagonist in wild-type mice. …”
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Crispant analysis in zebrafish as a tool for rapid functional screening of disease-causing genes for bone fragility by Sophie Debaenst, Tamara Jarayseh, Hanna De Saffel, Jan Willem Bek, Matthieu Boone, Ivan Josipovic, Pierre Kibleur, Ronald Y Kwon, Paul J Coucke, Andy Willaert

“…Six genes linked to severe recessive osteogenesis imperfecta (OI) and four associated with bone mineral density (BMD) from genome-wide association studies were analyzed using CRISPR/Cas9-based crispant screening in F0 mosaic founder zebrafish. …”
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TAp63γ is the primary isoform of TP63 for tumor suppression but not development by Xinbin Chen, Wenqiang Sun, Xiangmudong Kong, Xin Ming, Yanhong Zhang, Wensheng Yan, Shakur Mohibi, Mingyi Chen, Keith Mitchell, Jin Zhang

“…To explore this, we generated a mouse model in that exon 10’, a coding exon specific for p63γ, was deleted by CRISPR-cas9. We showed that mice deficient in p63γ are viable and futile, which is different from mice deficient in total TP63 or p63α. …”
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HP1 Promotes the Centromeric Localization of ATRX and Protects Cohesion by Interfering Wapl Activity in Mitosis by Erchen Zhang, Lei Peng, Kejia Yuan, Zexian Ding, Qi Yi

“…Methods: The clustered regularly interspaced short palindromic repeats with CRISPR-associated protein 9 (CRISPR-Cas9) system and overexpression approaches were employed alongside immunofluorescence to investigate the mechanism of ATRX localization at the centromere. …”
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Patient-derived response estimates from zero-passage organoids of luminal breast cancer by Róża K. Przanowska, Najwa Labban, Piotr Przanowski, Russell B. Hawes, Kristen A. Atkins, Shayna L. Showalter, Kevin A. Janes

“…Downstream mechanisms were tested genetically by lentiviral transduction of two complementary sgRNAs and Cas9 stabilization for the first week of organoid culture. …”
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Elite Alleles of EPE1 Identified via Genome-wide Association Studies Increase Panicle Elongation Length in Rice by Hao Sun, Qiqi Yao, Mei Hai, Tianhu Li, Jinghan Sun, Zhengbo Liu, Yang Ang, Yingying Zhao, Yanan Zhang, Xianping Cheng, Tao Huang, Yinping Chang, Mingyu Du, Erbao Liu

“…Compared with the short PELs of wild plants, mutant plants with Cas9-induced knockout (KO) of OsGA2ox9 exhibit long PELs. …”
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HPV16 <em>E6</em> Oncogene Contributes to Cancer Immune Evasion by Regulating PD-L1 Expression through a miR-143/HIF-1a Pathway by Georgios Konstantopoulos, Danai Leventakou, Despoina-Rozi Saltiel, Efthalia Zervoudi, Eirini Logotheti, Spyros Pettas, Korina Karagianni, Angeliki Daiou, Konstantinos E. Hatzistergos, Dimitra Dafou, Minas Arsenakis, Amanda Psyrri, Christine Kottaridi

“…The aim of this study was to investigate how these oncogenes contribute to the expression of immune system control molecules, using cell lines with integrated HPV16 genome, before and after knocking out <i>E6</i> viral gene using the CRISPR/Cas9 system, delivered with a lentiviral vector. …”
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Targeting and editing the second exon of bmp6 gene results in a silver carp with reduced intramuscular bones by Shuang Niu, Xiaohui Li, Cui Feng, Zewen Zhang, Hang Sha, Xiangzhong Luo, Guiwei Zou, Hongwei Liang

“…In this study, CRISPR/Cas9 technology was successfully used to target and disrupt exon 2 and exon 4 of bmp6 in silver carp, respectively, resulting in the generation of two bmp6-deficient chimeras, named chimera-E2 (CH-E2) and chimera-E4 (CH-E4). …”
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Tousled-like kinase loss confers PARP inhibitor resistance in BRCA1-mutated cancers by impeding non-homologous end joining repair by Min-ah Kim, Banseok Kim, Jihyeon Jeon, Jonghyun Lee, Hyeji Jang, Minjae Baek, Sang-Uk Seo, Dongkwan Shin, Anindya Dutta, Kyung Yong Lee

“…Loss-of-function experiments were performed in multiple human cancer cell lines using siRNA-mediated knockdown or CRISPR-Cas9 gene knockout. Cell viability assays were conducted to evaluate resistance to PARP inhibitors. …”
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Atlas of Fshr expression from novel reporter mice by Hongqian Chen, Hui-Qing Fang, Jin-Tao Liu, Shi-Yu Chang, Li-Ben Cheng, Ming-Xin Sun, Jian-Rui Feng, Ze-Min Liu, Yong-Hong Zhang, Clifford J Rosen, Peng Liu

“…To address this challenge, we developed Fshr-ZsGreen reporter mice under the control of Fshr endogenous promoter using CRISPR-Cas9. With this novel genetic tool, we provide a reliable readout of Fshr expression at single-cell resolution level in vivo and in real time. …”
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Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles by Julia Caspers, Andreas Ritter, Badi Bahrami, Samira Catharina Hoock, Susanne Roth, Alexandra Friemel, Franz Oswald, Frank Louwen, Nina-Naomi Kreis, Juping Yuan

“…As defective chromosome congression and segregation are the most remarkable features of cells depleted of RITA, we aimed to explore further potential related mechanisms, using various cellular and molecular techniques, including clustered regularly interspaced short palindromic repeats technique/deactivated CRISPR-associated protein 9 (CRISPR/dCas9), mass spectrometry, confocal microscopy, immunofluorescence, immunoprecipitation and Western blot analysis. …”
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Advancing the genetic engineering toolbox by combining AsCas12a knock-in mice with ultra-compact screening by Wei Jin, Yexuan Deng, John E. La Marca, Emily J. Lelliott, Sarah T. Diepstraten, Christina König, Lin Tai, Valentina Snetkova, Kristel M. Dorighi, Luke Hoberecht, Millicent G. Hedditch, Lauren Whelan, Geraldine Healey, Dan Fayle, Kieran Lau, Margaret A. Potts, Moore Z. Chen, Angus P. R. Johnston, Yang Liao, Wei Shi, Andrew J. Kueh, Benjamin Haley, Jean-Philippe Fortin, Marco J. Herold

“…Finally, we demonstrate CRISPR multiplexing via simultaneous gene knockout (via Cas12a) and activation (via dCas9-SAM) using primary T cells and fibroblasts. …”
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NOX4 modulates breast cancer progression through cancer cell metabolic reprogramming and CD8+ T cell antitumor activity by Yingying Xiong, Yiming Weng, Shan Zhu, Jian Qin, Jia Feng, Xiaopeng Jing, Chao Luo, Wei Gong, Rui Sun, Min Peng

“…Although NADPH oxidase 4 (NOX4) has been implicated in various oncogenic processes, its exact function in breast cancer progression, metabolic reprogramming, and immune modulation remains unclear.MethodsWe used murine 4T1 and EO771 breast cancer models to generate NOX4 knockout (KO) cell lines via CRISPR/Cas9. In vitro assays (cell proliferation, colony formation, wound healing, and Seahorse metabolic analyses) and in vivo orthotopic tumor studies assessed the impact of NOX4 loss. …”
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Silk Protein Gene Engineering and Its Applications: Recent Advances in Biomedicine Driven by Molecular Biotechnology by Ji X, Li Y, Wang J, Wang G, Ma B, Shi J, Cui C, Wang R

“…This review outlines the roles of major model organisms, such as silkworms and spiders, in silk protein production, and provides a detailed introduction to the applications of gene editing technologies (eg, CRISPR-Cas9), transgenic expression technologies, and synthetic biology techniques in silk protein genetic engineering. …”
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Cortisol regulates neonatal lung development via Smoothened by Shanshan Lu, Yifei Chen, Jiawen Song, Liangliang Ren, Jun Du, Donglai Shen, Jiayin Peng, Yao Yin, Xia Li, Yuqing Wang, Yan Gao, Siman Han, Yichang Jia, Yun Zhao, Yizheng Wang

“…To explore the effect of cortisol action on the SHH pathway on neonatal lung development, we generated a genetic mouse, in which leucine 116 (L112 in human) of SMO was mutated to alanine 116 (L116A, Smo a/a ) by the CRISPR-Cas9, based on sequence differences between human and mice. …”
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Genetic and pharmacological targeting of HINT2 promotes OXPHOS to alleviate inflammatory responses and cell necrosis in acute pancreatitis by Jiaqi Yao, Yuhong Jiang, Pengcheng Zhang, Yifan Miao, Xiajia Wu, Hang Lei, Zhijun Xie, Yong Tian, Xianlin Zhao, Juan Li, Lv Zhu, Meihua Wan, Wenfu Tang

“…Importantly, Emo upregulates the expression of HINT2 and OXPHOS complex proteins and enhances the interaction between HINT2 and MTCO2. Furthermore, CRISPR/Cas9-mediated HINT2 knockout significantly impaired the protective effects of Emo against AP-induced mitochondrial energy metabolism disorders, inflammatory responses, and acinar cell necrosis. …”
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