Article
Multidisciplinary Sciences
Yu Fu, Zhenglin Zhu, Geng Meng, Rijun Zhang, Yueping Zhang
Summary: Post-translational modifications of histone proteins play a crucial role in gene expression and cell fate decisions. Developing efficient methods for introducing mutations to histones is vital for studying these modifications. By utilizing a CRISPR-Cas9 based system in yeast cells, researchers successfully generated multiple histone mutants, presenting a valuable tool for the field of epigenetics.
SCIENTIFIC REPORTS
(2021)
Article
Multidisciplinary Sciences
Arvind Sharma, Michael N. Pham, Jeremiah B. Reyes, Randeep Chana, Won C. Yim, Chan C. Heu, Donghun Kim, Duverney Chaverra-Rodriguez, Jason L. Rasgon, Robert A. Harrell, Andrew B. Nuss, Monika Gulia-Nuss
Summary: Ticks have the ability to transmit various debilitating pathogens, but research on ticks has been lacking compared to other arthropod vectors due to challenges in applying genetic and molecular tools. This study reports a successful embryo injection protocol for the black-legged tick and demonstrates the use of CRISPR-Cas9 for genome editing. The results provide innovative tools for tick research.
Article
Plant Sciences
Bicko Steve Juma, Asunta Mukami, Cecilia Mweu, Mathew Piero Ngugi, Wilton Mbinda
Summary: This study successfully developed cassava plants with lower levels of cyanogenic glycoside and cyanide content through CRISPR/Cas9-mediated targeted mutagenesis. This provides an alternative approach for breeding non-toxic cassava varieties.
FRONTIERS IN PLANT SCIENCE
(2022)
Article
Fisheries
Qian Li, Hong Yu, Qi Li
Summary: Genome editing using CRISPR/Cas9 methodology holds great potential for improving aquaculture, but the screening and identification of successfully edited individuals remains challenging. In this study, we developed and optimized a high-resolution melting analysis (HRM) assay to rapidly screen and identify CRISPR/Cas9-edited oysters. The HRM assay showed sensitivity in detecting temperature shifts and successfully identified oysters with indel mutations. This reproducible, sensitive, and efficient method has the potential to accelerate genetic improvement in aquaculture.
Review
Biochemistry & Molecular Biology
Caixia Gao
Summary: Genome editing has the potential to revolutionize plant breeding and address the challenge of feeding a growing population in the face of climate change. Newly developed techniques and strategies offer significant opportunities for plant improvement and crop production, but challenges must be overcome to realize the full potential of this technology.
Article
Biochemistry & Molecular Biology
Ramona Gruetzner, Patrick Martin, Claudia Horn, Samuel Mortensen, Erin J. Cram, Carolyn W. T. Lee-Parsons, Johannes Stuttmann, Sylvestre Marillonnet
Summary: The recent discovery of the mode of action of the CRISPR/Cas9 system has provided biologists with a useful tool for generating site-specific mutations in genes of interest. In this study, optimizing the efficiency of the Cas9 nuclease in generating mutations in target genes by investigating its nucleotide and/or amino acid sequence features led to the finding that adding introns to the Cas9 coding sequence significantly improved editing efficiency. The intronized Cas9 gene was shown to be effective in generating mutant phenotypes in various plants, highlighting the importance of sequence modifications for enhancing CRISPR/Cas9 genome editing.
PLANT COMMUNICATIONS
(2021)
Article
Plant Sciences
Chong Ren, Yanfei Liu, Yuchen Guo, Wei Duan, Peige Fan, Shaohua Li, Zhenchang Liang
Summary: This study identified multiple effective promoters in grapevine and achieved multiplex genome editing by optimizing the CRISPR/Cas9 system, leading to increased editing efficiency. Simultaneous editing of TMT1 and TMT2 genes resulted in reduced sugar levels in grapes, highlighting the role of these genes in sugar accumulation. The findings provide valuable materials for the optimization of CRISPR/Cas9 system and expand the toolbox of grape genome editing for basic research and molecular breeding.
HORTICULTURE RESEARCH
(2021)
Article
Plant Sciences
Mengxi Bi, Zhijun Wang, Keyan Cheng, Yiqing Cui, Yi He, Jian Ma, Mingfang Qi
Summary: This study successfully applied the CRISPR/Cas9 large-scale screening strategy in tomato and constructed a CRISPR/Cas9 mutant library. The study found that homozygous and biallelic mutations occurred at higher frequencies in the mutant population. Additionally, a small-scale CRISPR/Cas9 library enhanced the efficacy of single sgRNA recognition in positive plants.
PLANT PHYSIOLOGY AND BIOCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Chaya T. L. Yuen, Dawn G. L. Thean, Becky K. C. Chan, Peng Zhou, Cynthia C. S. Kwok, Hoi Yee Chu, Maggie S. H. Cheung, Bei Wang, Yee Man Chan, Silvia Y. L. Mak, Anskar Y. Leung, Gigi C. G. Choi, Zongli Zheng, Alan S. L. Wong
Summary: Researchers have enhanced editing accuracy and reduced off-target edits by re-engineering KKH-SaCas9 with a Y239H mutation and a set of additional mutations, demonstrating the feasibility of multi-domain combinatorial mutagenesis to optimize editing fidelity.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Plant Sciences
Bhuvan Pathak, Chandan Maurya, Maria C. Faria, Zahra Alizada, Soumen Nandy, Shan Zhao, Muhammed K. Jamsheer, Vibha Srivastava
Summary: This study demonstrates the efficiency of CRISPR/Cas9 in creating mutations in essential genes in rice, although the mutagenesis rates vary among different genes. Additionally, the study shows that the generated mutants exhibit phenotypes such as sterility or early senescence.
Article
Plant Sciences
Xavier Herman, Johann Far, Adeline Courtoy, Laurent Bouhon, Loic Quinton, Edwin De Pauw, Francois Chaumont, Catherine Navarre
Summary: By using CRISPR/Cas9 technology to edit two N-acetylglucosaminyltransferase I (GnTI) genes, a Nicotiana tabacum Bright Yellow-2 (BY-2) cell line that produces glycoproteins devoid of complex N-glycans was successfully obtained.
FRONTIERS IN PLANT SCIENCE
(2021)
Article
Multidisciplinary Sciences
Vanessa Chenouard, Isabelle Leray, Laurent Tesson, Severine Remy, Alasdair Allan, Daniel Archer, Adam Caulder, Agnes Fortun, Karine Bernardeau, Yacine Cherifi, Lydia Teboul, Laurent David, Ignacio Anegon
Summary: CRISPR-Cas9 cleavage efficacy and accuracy are influenced by the optimal formation of the ribonucleoprotein (RNP) complex. The study shows that an equimolar ratio of Cas9 and guide RNA (gRNA) is optimal for RNP complex formation. Increasing the Cas9/gRNA ratio does not improve knock-in (KI) efficiency, and excess gRNA leads to a decrease in point mutation KI efficiency and an increase in large deletions.
Review
Chemistry, Multidisciplinary
Li Duan, Limei Xu, Xiao Xu, Zhuan Qin, Xiaoying Zhou, Yin Xiao, Yujie Liang, Jiang Xia
Summary: Gene vectors deliver genetic materials or gene editing devices into cells for therapeutic protein production or gene correction, but barriers like cell membrane and degradation necessitate carriers for clinical gene therapy. Exosomes, engineered to encapsulate and deliver nucleic acids, and their fusion with liposomes increase loading capacity and maintain targeting capability, showing promise for future gene delivery applications in therapy.
Article
Biotechnology & Applied Microbiology
Hai-yue Bao, Hui-jie Li, Yong-yong Zhang, Andreas Bechthold, Xiao-ping Yu, Zheng Ma
Summary: In this study, a transposon mutagenesis strategy was employed to identify an unknown gene involved in rimocidin biosynthesis in Streptomyces rimosus M527. The results showed that the genes rrt and hyp have a positive and negative impact on rimocidin production, respectively, without affecting cell growth.
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
(2023)
Article
Immunology
Run Fang, Qifei Jiang, Yukun Guan, Pengfei Gao, Rui Zhang, Zhen Zhao, Zhengfan Jiang
Summary: The biosynthesis of sulfated glycosaminoglycans (sGAGs) in the Golgi apparatus is crucial for the polymerization and activation of the STING receptor. sGAGs promote STING polymerization through luminal, positively charged, polar residues, and the chain length and sulfation of sGAGs directly affect STING activation. The translocation from the endoplasmic reticulum to the Golgi apparatus is necessary for STING activation.
Article
Biochemistry & Molecular Biology
Bozena Szulc, Yelyzaveta Zadorozhna, Mariusz Olczak, Wojciech Wiertelak, Dorota Maszczak-Seneczko
Summary: This study examined how specific mutations in the SLC35A1 gene affect the protein's properties, revealing that CDG-causing mutations do not affect protein localization in the Golgi apparatus but do impact CST functionality, especially when T156R and E196K mutations are combined. Additionally, differences in the ability of CST variants to form dimers were uncovered, improving understanding of SLC35A1-CDG cases.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Review
Biochemistry & Molecular Biology
Dorota Maszczak-Seneczko, Maciej Wiktor, Edyta Skurska, Wojciech Wiertelak, Mariusz Olczak
Summary: This review presents a comprehensive overview of the import of nucleotide sugars into the mammalian Golgi, including experimental approaches to study NS transport, various aspects of knowledge concerning NSTs, and the pathologies related to NSTs' defective function. The outcomes of research using mammalian cell-based models are also discussed.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Edyta Skurska, Bozena Szulc, Dorota Maszczak-Seneczko, Maciej Wiktor, Wojciech Wiertelak, Aleksandra Makowiecka, Mariusz Olczak
Summary: Mutations in the SLC35C1 gene can cause leukocyte adhesion deficiency II, but the improvement of fucosylation in patients suggests the existence of an SLC35C1-independent route of GDP-fucose transport.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2022)
Article
Cell Biology
Magdalena Kot, Ewa Mazurkiewicz, Maciej Wiktor, Wojciech Wiertelak, Antonina Joanna Mazur, Andrei Rahalevich, Mariusz Olczak, Dorota Maszczak-Seneczko
Summary: SLC35A2 deficiency promotes epithelial-to-mesenchymal transition (EMT), leading to cell morphological changes, impaired cell-cell adhesion, altered gene expression, and Golgi reorganization.
Article
Biochemistry & Molecular Biology
Wojciech Wiertelak, Karolina Chabowska, Bozena Szulc, Yelyzaveta Zadorozhna, Mariusz Olczak, Dorota Maszczak-Seneczko
Summary: Nucleotide sugar transporters (NSTs) are crucial for providing glycosylation enzymes with substrates. Although their interactions with glycosyltransferases involved in N-glycosylation have been studied, their interactions with enzymes involved in mucin-type O-glycan biosynthesis remain unclear. This study reveals the association between UDP-galactose transporter (UGT) and core 1 & beta;-1,3-galactosyltransferase 1 (C1GalT1) in the O-glycosylation pathway, providing the first example of an NST interacting with an enzyme in this pathway. The study also suggests that NSTs may play a role in stabilizing interaction partners and facilitating target localization in cells.
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
(2023)
Article
Biochemistry & Molecular Biology
Wojciech Wiertelak, Artem Pavlovskyi, Dorota Maszczak-Seneczko, Bozena Szulc, Mariusz Olczak
Summary: SLC35A2 and SLC35A3 are homologous proteins with postulated nucleotide sugar transporting activities. The UDP-GlcNAc transporting activity initially attributed to SLC35A3 has recently been put into question. Constructed hybrid proteins (SLC35A2-SLC35A3 and SLC35A3-SLC35A2) were able to restore glycosylation in a SLC35A2/SLC35A3 double knockout cell line, indicating a balanced cooperation between SLC35A2 and SLC35A3 is required for proper glycosylation.
