Latest Publications on Biotechnology and CRISPR
This is our latest selection of worldwide publications on Biotechnology and CRISPR, between many scientific online journals, classified and focused on CRISPR, gene editing, gRNA, PAM sequence, DNA cleavage, Cas9 protein, genome editing, gene insertion and gene knockout.
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Construction of a TAT-Cas9-EGFP Site-Specific Integration Eukaryotic Cell Line Using Efficient PEG10 Modification
On 2025-02-04 by Shiyu Qi, Yibo Wang, Zhimei Liu, Sujun Wu, Yue Zhao, Yan Li, Shoulong Deng, Kun Yu, Zhengxing Lian @MDPI
Abstract: The CRISPR/Cas9 system enables precise and efficient modification of eukaryotic genomes. Among its various applications, homology-directed repair (HDR) mediated knock-in (KI) is crucial for creating human disease models, gene therapy, and agricultural genetic enhancements. Despite its potential, HDR-mediated knock-in efficiency remains relatively low. This study investigated the impact of 5&prime; end PEG10 modification on site-specific integration of the target gene. The HEK293 cell lin[...]
AI summary: Construction of TAT-Cas9-EGFP cell line with PEG10 modification. Investigation of HDR-mediated knock-in efficiency. Utilization of HEK293 cell line for stable protein expression. Increase in knock-in efficiency with PEG10 modification.
CRISPR/Cas9, Eukaryotic Cell Line, PEG10 Modification, Site-Specific Integration, HDR-mediated knock-in, HEK293 cell line, AAVS1 locus
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ctdsp2 Knockout Induces Zebrafish Craniofacial Dysplasia via p53 Signaling Activation
On 2025-02-03 by Xin Xia, Wenjie Song, Fuyu Zhang, Yue Fan, Bo Zhang, Xiaowei Chen @MDPI
Abstract: Hemifacial microsomia (HFM) is a rare congenital craniofacial deformity that significantly impacts the appearance and hearing. The genetic etiology of HFM remains largely unknown, although genetic factors are considered to be primary contributors. We previously identified CTDSP2 as a potential causative gene in HFM cases. Utilizing CRISPR/Cas9, we knocked out ctdsp2 in zebrafish and analyzed the spatiotemporal expression of ctdsp2 and neural crest cell (NCC) markers through in situ hybridization[...]
AI summary: Knockout of ctdsp2 in zebrafish induces craniofacial dysplasia. Abnormalities in NCC apoptosis and proliferation observed. RNA-Seq analysis reveals upregulation of p53 signaling genes. ctdsp2 knockout disrupts chondrocyte differentiation and inhibits NCC proliferation.
ctdsp2, Knockout, Zebrafish, Craniofacial Dysplasia, p53 Signaling, HFM, Genetic factors
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Subgenomic RNA Detection in SARS-CoV-2 Assessing Replication and Inactivation Through Serial Passages, RT-qPCR, and Electron Microscopy
On 2025-02-01 by Talita da Silva França, Juliana Fernandes Amorim da Silva, Gabriella Christine Neves da Silva, Barbara Oliveira dos Santos, Stephanie Almeida Silva, José Henrique Resende Linhares, Marcos Alexandre Nunes da Silva, Debora Ferreira Barreto-Vieira, Vanessa Salete de Paula, Liliane Monteiro de Morais, Renata Tourinho Santos, Gisela Freitas Trindade @MDPI
Abstract: Subgenomic RNAs (sgRNAs) are potential markers of active SARS-CoV-2 replication, serving as templates for the synthesis of structural and accessory proteins in infectious viral particles. This study aimed to use RT-qPCR to quantify sgRNA and negative RNA intermediates, assessing viral replication in virus samples inactivated by &beta;-propiolactone (&beta;PL). Inactivated viruses subjected to five blind serial passages (BSs) were amplified by RT-qPCR using primers to target the e[...]
AI summary: Subgenomic RNA detection in SARS-CoV-2 assessed through RT-qPCR and electron microscopy. Inactivation of viral samples by β-propiolactone confirmed by serial passages. Replication kinetics showed exponential increase in viral titer, supporting sgRNA as reliable marker for active replication. Transmission electron microscopy confirmed presence of viral particles in infected cells.
subgenomic RNA, SARS-CoV-2, RT-qPCR, viral replication, inactivation, electron microscopy, diagnostic strategies
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Current Understanding and Methodological Challenges
On 2025-01-30 by Polina N. Kamzeeva, Vera A. Alferova, Vladimir A. Korshun, Anna M. Varizhuk, Andrey V. Aralov @MDPI
Abstract: RNA G-quadruplexes (rG4s) in 5&prime;-UTRs represent complex regulatory elements capable of both inhibiting and activating mRNA translation through diverse mechanisms in eukaryotes. This review analyzes the evolution of our understanding of 5&prime;-UTR rG4-mediated translation regulation, from early discoveries of simple translation inhibitors to the current recognition of their multifaceted regulatory roles. We discuss canonical and non-canonical rG4 structures, their interacti[...]
AI summary: Evolution of understanding of RNA G-quadruplexes in translation regulation, diverse regulatory roles, interactions with regulatory proteins, synergistic effects with uORFs, methodological challenges in studying RNA G-quadruplexes, potential therapeutic targets.
RNA G-quadruplexes, translation regulation, 5&prime;-UTRs, helicases, IRES-mediated translation, methodological challenges, neurodegenerative diseases
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p21 Promoter Methylation Is Vital for the Anticancer Activity of Withaferin A
On 2025-01-30 by Andrew Brane, Madeline Sutko, Trygve O. Tollefsbol @MDPI
Abstract: Breast cancer (BC) is a widespread malignancy that affects the lives of millions of women each year, and its resulting financial and healthcare hardships cannot be overstated. These issues, in combination with side effects and obstacles associated with the current standard of care, generate considerable interest in new potential targets for treatment as well as means for BC prevention. One potential preventive compound is Withaferin A (WFA), a traditional medicinal compound found in winter cherr[...]
AI summary: p21 promoter methylation is essential for the anticancer activity of Withaferin A in breast cancer. Financial and healthcare hardships of BC drive interest in new treatment targets. WFA shows promise as a preventive compound, acting through epigenetic mechanisms. Targeting specific gene promoters, such as p21, p53, and CCND1, can modulate the anticancer effects of WFA.
p21 Promoter, Methylation, Withaferin A, Breast Cancer, Epigenome, Anticancer Activity, Methylome
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Perspectives of Genome Editing Mediated Haploid Inducer Systems in Legumes
On 2025-01-29 by Yiqian Liu, Musazade Elshan, Geng Li, Xiao Han, Xiao Chen, Xianzhong Feng @MDPI
Abstract: Genome editing-mediated haploid inducer systems (HISs) present a promising strategy for enhancing breeding efficiency in legume crops, which are vital for sustainable agriculture due to their nutritional benefits and ability to fix nitrogen. Traditional legume breeding is often slow and complicated by the complexity of legumes&rsquo; genomes and the challenges associated with tissue culture. Recent advancements have broadened the applicability of HISs in legume crops, facilitating a redu[...]
AI summary: Perspectives on genome editing in legume breeding. HISs offer a promising strategy for enhancing efficiency. Integration of genome editing technology accelerates trait development. Additional research needed to optimize systems for sustainability.
genome editing, haploid inducer systems, legumes, breeding efficiency, sustainable agriculture, genetic modifications, homozygous lines
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