Development of insect cell line using CRISPR technology

Nweke, Ekene Emmanuel; Raj, Deepak

doi:10.1016/bs.pmbts.2021.01.003

Cited by 4 publications

(3 citation statements)

References 65 publications

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“…In this cell line, the inactivation of CDKN2A at two loci induced immortalization, but also maintained the characteristics of normal cells, including a normal p53 response [ 56 ] . CRISPR-Cas9 has also been used to edit several existing insect cell lines, including insect cell lines Sf-21 and S2R+ [ 78 , 79 ] . Despite great potential, CRISPR-Cas9 has not yet been employed to produce a novel insect cell line.…”

Section: Producing Immortalized Insect Gut Cell Linesmentioning

confidence: 99%

The untapped potential of cell culture in disentangling insect-microbial relationships

Macpherson,

Daisley,

Mallory

et al. 2024

Microbiome Res Rep

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Cell culture is a powerful technique for the investigation of molecular mechanisms fundamental to health and disease in a diverse array of organisms. Cell lines offer several advantages, namely their simplistic approach and high degree of reproducibility. One field where cell culture has proven particularly useful is the study of the microbiome, where cell culture has led to the illumination of microbial influences on host immunity, nutrition, and physiology. Thus far, researchers have focused cell culture work predominantly on humans, but the growing field of insect microbiome research stands to benefit greatly from its application. Insects constitute one of Earth’s most diverse and ancient life forms and, just as with humans, possess microbiomes with great significance to their health. Insects, which play critical roles in supporting food security and ecological stability, are facing increasing threats from agricultural intensification, climate change, and pesticide use. As the microbiome is closely tied to host health, gaining a more robust understanding is of increasing importance. In this review, we assert that the cultivation and utilization of insect gut cell lines in microbiome research will bridge critical knowledge gaps essential for informing insect management practices in a world under pressure.

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Section: Producing Immortalized Insect Gut Cell Linesmentioning

confidence: 99%

The untapped potential of cell culture in disentangling insect-microbial relationships

Macpherson,

Daisley,

Mallory

et al. 2024

Microbiome Res Rep

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“…Of note, a technique for CRISPR/Cas9 gene editing in Sf9 insect cells was first performed by Donald L. Jarvis and colleagues [ 126 ]. The team used the CRISPR-Cas9 vector as a platform to produce recombinant glycoproteins [ 127 ]. They also evaluated the potential for site-specific gene editing in baculovirus insect cell system (BICS) host insect cell lines by using several types of insect cell U6 promoters upstream of gene cassette to create CRISPR-Cas9 expression vector in Sf9 and High Five cells.…”

Section: The Future Of Crispr/cas9 To Complex Glycosylation In Bicsmentioning

confidence: 99%

“…The generic CRISPR-Cas9 vectors with DmU6 or BmU6-2 promoters did not allow CRISPR-Cas9 gene editing in Sf9 cells. However, the ability to manufacture indels was confirmed by sequencing using the CRISPR-Cas9 vector that has a SfU6-3 promoter (rather than DmU6 or BmU6-2) [ 127 ].…”

Section: The Future Of Crispr/cas9 To Complex Glycosylation In Bicsmentioning

confidence: 99%

Advances in CRISPR-Cas9 for the Baculovirus Vector System: A Systematic Review

Sari-Ak

Alomari

Al-Shomali

et al. 2022

Viruses

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The baculovirus expression vector systems (BEVS) have been widely used for the recombinant production of proteins in insect cells and with high insert capacity. However, baculovirus does not replicate in mammalian cells; thus, the BacMam system, a heterogenous expression system that can infect certain mammalian cells, was developed. Since then, the BacMam system has enabled transgene expression via mammalian-specific promoters in human cells, and later, the MultiBacMam system enabled multi-protein expression in mammalian cells. In this review, we will cover the continual development of the BEVS in combination with CRPISPR-Cas technologies to drive genome-editing in mammalian cells. Additionally, we highlight the use of CRISPR-Cas in glycoengineering to potentially produce a new class of glycoprotein medicines in insect cells. Moreover, we anticipate CRISPR-Cas9 to play a crucial role in the development of protein expression systems, gene therapy, and advancing genome engineering applications in the future.

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Serum‐free medium for recombinant protein expression in insect cells

Geng,

Zou,

Bai

et al. 2024

Biotech and App Biochem

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The baculovirus expression vector system (BEVS) has been widely used to produce recombinant proteins because of several advantages, such as eukaryotic post‐translational modifications similar to those in mammalian cells, high expression levels and safety, and large gene capacity. Usually, insect cell culture requires 5%‒10% fetal bovine serum, which has many adverse effects, including high cost, heterogeneity between batches, complex composition, and pollution risks. Therefore, serum‐free medium (SFM) is indispensable for the production of recombinant proteins in insect cell culture. Here, the most commonly used insect cell lines and three insect cell media, namely basic medium, SFM, and chemically defined medium, are summarized. The basic components of insect cell SFM are similar to those of other cells but contain special components. The components, functions, and issues of different SFM used for insect cell culture are reviewed. In recent years, some special additives have been demonstrated to increase recombinant protein expression yield and quality in BEVS, and the functions and possible mechanisms of small‐molecule additives are reviewed herein. Finally, future perspectives of SFM used in BEVS for recombinant protein production are discussed.

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Development of insect cell line using CRISPR technology

Cited by 4 publications

References 65 publications

The untapped potential of cell culture in disentangling insect-microbial relationships

The untapped potential of cell culture in disentangling insect-microbial relationships

Advances in CRISPR-Cas9 for the Baculovirus Vector System: A Systematic Review

Serum‐free medium for recombinant protein expression in insect cells

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