Electroporation Mediated DNA Transformation of Filamentous Fungi

Chakraborty, B. N.

doi:10.1007/978-3-319-10142-2_6

Cited by 4 publications

(2 citation statements)

References 87 publications

(64 reference statements)

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“…Electroporation is very simple and less time‐consuming, especially for spore‐producing species for which PMT is difficult [40]. It uses a high‐voltage electric current that creates permeability in the membrane, making the membrane prone to receive foreign genetic materials.…”

Section: Traditional Genetic Transformation Techniques In Filamentousmentioning

confidence: 99%

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

Ullah

Lin

Xie

et al. 2020

Biotech and App Biochem

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Filamentous fungi have several industrial, environmental, and medical applications. However, they are rarely utilized owing to the limited availability of full‐genome sequences and genetic manipulation tools. Since the recent discovery of the full‐genome sequences for certain industrially important filamentous fungi, CRISPR/Cas9 technology has drawn attention for the efficient development of engineered strains of filamentous fungi. CRISPR/Cas9 genome editing has been successfully applied to diverse filamentous fungi. In this review, we briefly discuss the use of common genetic transformation techniques as well as CRISPR/Cas9‐based systems in filamentous fungi. Furthermore, we describe potential limitations and challenges in the practical application of genome engineering of filamentous fungi. Finally, we provide suggestions and highlight future research prospects in the area.

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Section: Traditional Genetic Transformation Techniques In Filamentousmentioning

confidence: 99%

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

Ullah

Lin

Xie

et al. 2020

Biotech and App Biochem

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“…This method has shown its capacity in introducing DNA plasmids or other vectors in various types of applications, ranging from cells to yeast and E-coli (such as ref. 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 ), from in vitro studies to in vivo tissues 11 12 13 14 15 16 17 . Because of the advantages of broad applicability, rapidity, technical simplicity and avoidance of using toxic chemicals, electroporation is becoming a promising tool in the research of gene therapy and DNA vaccination.…”

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confidence: 99%

Electroporation on microchips: the harmful effects of pH changes and scaling down

Zhao

et al. 2015

Sci Rep

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Electroporation has been widely used in delivering foreign biomolecules into cells, but there is still much room for improvement, such as cell viability and integrity. In this manuscript, we investigate the distribution and the toxicity of pH changes during electroporation, which significantly decreases cell viability. A localized pH gradient forms between anode and cathode leading to a localized distribution of cell death near the electrodes, especially cathodes. The toxicity of hydroxyl ions is severe and acute due to their effect in the decomposition of phospholipid bilayer membrane. On the other hand, the electric field used for electroporation aggravates the toxicity of hydroxyl because the electropermeabilization of cell membrane makes bilayer structure more loosen and vulnerable. We also investigate the side effects during scaling down the size of electrodes in electroporation microchips. Higher percentage of cells is damaged when the size of electrodes is smaller. At last, we propose an effective strategy to constrain the change of pH by modifying the composition of electroporation buffer. The modified buffer decreases the changes of pH, thus enables high cell viability even when the electric pulse duration exceeds several milliseconds. This ability has potential advantage in some applications that require long-time electric pulse stimulation.

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Metabolic Engineering of Fungal Strains for Efficient Production of Cellulolytic Enzymes

Zhao

Zhang

et al. 2018

Fungal Cellulolytic Enzymes

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Electroporation Mediated DNA Transformation of Filamentous Fungi

Cited by 4 publications

References 87 publications

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

CRISPR/Cas9‐based genome engineering: A new breakthrough in the genetic manipulation of filamentous fungi

Electroporation on microchips: the harmful effects of pH changes and scaling down

Metabolic Engineering of Fungal Strains for Efficient Production of Cellulolytic Enzymes

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