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Construction and characterization of lpxC deletion strain based on CRISPR/Cas9 in Acinetobacter baumannii
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Zong-ti SUN1, You-wen ZHANG1, 2, Hai-bin LI1, Xiu-kun WANG1, 2, Jie YU1, Jin-ru XIE1, Peng-bo PANG1, Xin-xin HU1, 2, Tong-ying NIE1, 2, Xi LU1, 2, Jing PANG1, 2, Lei HOU3, Xin-yi YANG1, 2, Cong-ran LI1, 2, Lang SUN1, 2, *, Xue-fu YOU1, 2, *
Acta Pharmaceutica Sinica | 2024, 59(5) : 1286 - 1294
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Acta Pharmaceutica Sinica | 2024, 59(5): 1286-1294
• Original Articles •
Construction and characterization of lpxC deletion strain based on CRISPR/Cas9 in Acinetobacter baumannii
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Zong-ti SUN1, You-wen ZHANG1, 2, Hai-bin LI1, Xiu-kun WANG1, 2, Jie YU1, Jin-ru XIE1, Peng-bo PANG1, Xin-xin HU1, 2, Tong-ying NIE1, 2, Xi LU1, 2, Jing PANG1, 2, Lei HOU3, Xin-yi YANG1, 2, Cong-ran LI1, 2, Lang SUN1, 2, *, Xue-fu YOU1, 2, *
Affiliations
  • 1. Beijing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
  • 2. Division for Medicinal Microorganisms Related Strains, CAMS Collection Center of Pathogenic Microorganisms, Beijing 100050, China
  • 3. Experimental Animal Center, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
Published: 2024-05-12 doi: 10.16438/j.0513-4870.2024-0044
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Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D9). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

Acinetobacter baumannii  /  gene editing  /  CRISPR/Cas9 system  /  lpxC gene  /  polymyxin  /  resistance  /  lipopolysaccharide
Zong-ti SUN, You-wen ZHANG, Hai-bin LI, Xiu-kun WANG, Jie YU, Jin-ru XIE, Peng-bo PANG, Xin-xin HU, Tong-ying NIE, Xi LU, Jing PANG, Lei HOU, Xin-yi YANG, Cong-ran LI, Lang SUN, Xue-fu YOU. Construction and characterization of lpxC deletion strain based on CRISPR/Cas9 in Acinetobacter baumannii[J]. Acta Pharmaceutica Sinica, 2024 , 59 (5) : 1286 -1294 . DOI: 10.16438/j.0513-4870.2024-0044
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Article Info
doi: 10.16438/j.0513-4870.2024-0044
  • Receive Date：2024-01-16
  • Online Date：2025-11-27
  • Published：2024-05-12
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History
  • Received：2024-01-16
  • Revised：2024-02-21
Funding
Affiliations
    1. Beijing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
    2. Division for Medicinal Microorganisms Related Strains, CAMS Collection Center of Pathogenic Microorganisms, Beijing 100050, China
    3. Experimental Animal Center, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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