Genome centric engineering using ZFNs, TALENs and CRISPR-Cas9 systems for trait improvement and disease control in Animals

Wani, Atif Khurshid; Akhtar, Nahid; Singh, Reena; Prakash, Ajit; Raza, Sayed Haidar Abbas; Cavalu, Simona; Chopra, Chirag; Madkour, Mahmoud; Elolimy, Ahmed A.; Hashem, Nesrein M.

doi:10.1007/s11259-022-09967-8

Cited by 21 publications

(11 citation statements)

References 172 publications

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“…2-uorofucose and 5-alkynylfucose derivatives were utilized as fucosylation inhibitors to produce afucosylated antibodies. However, this approach was not recommended because of the toxicity to cells [21], then gene knockout techniques were applied instead, including zinc-nger nucleases (ZFNs) [22,23], transcription activator-like effector nucleases (TALENs) [24], and CRISPR/Cas9 [25][26][27]. The FokI is an arti cial endonuclease composed of a zinc nger protein and the cleavage domain [28], and the TALEN is produced by the fusion of a transcription activator-like effector and the nuclease catalytic domain of FokI.…”

Section: Discussionmentioning

confidence: 99%

Establishment of FUT8 gene knockout CHO cell line with stable expression of monoclonal antibody

Gao,

Zhang,

et al. 2023

Preprint

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Background: Afucosylated antibodies have been become increasingly popular in clinical trials for therapeutic purposes owing to the improved efficacy compared to the fucosylated counterparts. The elimination of fucose on the heavy chain of an antibody can enhance the activity of antibody-dependent cellular cytotoxicity (ADCC). However, antibodies produced by Chinese hamster ovary (CHO) cells are heavily fucosylated, and the modification reduces the activity of ADCC. FUT8,GMD and FX encoding major fucose modification enzymes, which were knockout respectively in CHO cells using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9). These gene knockout cell lines were evaluated for cell growth, antibody yield, N-glycan profiles and ADCC activities. Results: The results showed that the gene knockout cell lines exhibited morphology and growth kinetics similar to those of the wild CHO cells, and produced completely afucosylated recombinant antibodies. Among them, the ADCC activity of cell pool and monoclonal cell line with FUT8 gene knocked out (FUT8KO) were increased 2-2.5-fold comparing to conventional fucosylated antibodies. Conclusions: Our results indicated that FUT8KO monoclonal clone cell lines (#08E9, 05E9, 20D8, 12C4, 01E9, and 01F4) were ideal hosts for stable production of completely afucosylated high-ADCC activity antibodies, with considerable therapeutic quality and efficacy.

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Section: Discussionmentioning

confidence: 99%

Establishment of FUT8 gene knockout CHO cell line with stable expression of monoclonal antibody

Gao,

Zhang,

et al. 2023

Preprint

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“…Scientists can then insert, delete, or replace the cut DNA sequence with a desired sequence (Mehra and Kumar, 2022;Rasheed et al, 2022;Khiabani et al, 2023;Li et al, 2023), (b) Zinc Finger Nucleases (ZFNs): Zinc Finger Nucleases are engineered proteins that can bind to specific DNA sequences and cut the DNA at that location. Like CRISPR-Cas9, ZFNs can be used to add, delete or modify genes (Mehra and Kumar, 2022;Wani et al, 2023a), (c) TALENs: Transcription Activator-Like Effector Nucleases (TALENs) are another type of engineered proteins that can bind to specific DNA sequences and cut the DNA at that location, and TALENs work by using a DNA-binding domain that can be programmed to recognize a specific DNA sequence (Mehra and Kumar, 2022;Kumar and Kues, 2023;Wani et al, 2023a), (d) Homologous Recombination: Homologous Recombination is a technique that uses a DNA template to repair a broken DNA strand. This technique is often used to introduce specific mutations or changes to a DNA sequence (Ranjitha et al, 2022;Park, 2023).…”

Section: Genome Editing Techniquesmentioning

confidence: 99%

“… Editing the genes in the desired animals: Once the appropriate gene-editing technique has been chosen, the next step is to edit the genes in the desired animals. This could involve introducing new genes, deleting or disabling existing genes, or modifying the expression of genes (Mushtaq and Molla, 2023;Wani et al, 2023b).…”

Section: Black Sea Journal Of Agriculturementioning

confidence: 99%

Utilization of Genome Editing for Livestock Resilience in Changing Environment

Ngeno

2023

Black Sea Journal of Agriculture

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Climate change poses a significant threat to livestock production systems, including changes in temperature and rainfall patterns, increased frequency of extreme weather events, and the spread of diseases. The use of genome editing technologies presents a potential solution to mitigate the impacts of climate change on livestock. This paper reviewed the prospects of utilizing genome editing in mitigating the impact of climate change in livestock. Applications of genome editing in development of heat-tolerant, and disease-resistant as well as animals with improved feed and water use efficiency and reduced methane emissions are explored. Additionally, a potential breeding program for gene edited animals is proposed. There are several different genome editing techniques that can be used in livestock breeding, including CRISPR/Cas9, TALENs, and zinc-finger nucleases. These techniques involve introducing specific changes to the animal's genome, such as deleting or replacing genes, or introducing new ones. The technology has enormous potential for improving livestock breeding, as it allows for the creation of animals with desirable traits in a much shorter time frame than traditional breeding methods. Generally, it may take years or even decades to breed an animal with a specific trait using traditional breeding methods, whereas genome editing can achieve the same result in just a few generations. Genome editing can be used to mitigate the impact of climate change on livestock production by reducing the methane emissions by improving the efficiency of feed conversion and modifying the genes responsible for methane production. Technology can be utilized to improve livestock feeds by modifying genes involved in plant growth, development, and nutrient use. This lead to the creation of forages that are high yielding, more nutritious and better adapted to diverse production environments. Genome editing allows development of animals that are more resistant to diseases, which can help reduce the need for antibiotics and other treatments. This is particularly important given the growing problem of antibiotic resistance, which is a major concern in both human and animal health. Genome editing has the potential of developing animals that are thermo-tolerant, as well as animals with improved feed and water use efficiency. The proposed breeding program for gene-edited animals will ensure that the animals produced are healthy, genetically diverse, and meet the desired traits. In terms of ethical concerns, policies for genome editing ought consider the potential for unintended consequences or the creation of animals with characteristics that are viewed as undesirable or unethical Overall, genome editing technology has the potential to revolutionize livestock production and contribute to the global effort to mitigate the impact of climate change.

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“…The security challenges posed by genome editing, which allows the alteration of genes, also pose a security challenge for law enforcement agencies. Various guidelines have been established by regulatory authorities worldwide to keep the development of genome editing technologies in control [ 251 , 252 ].…”

Section: Biosensors For Detection Of Biological- and Chemical-warfare...mentioning

confidence: 99%

Advancing biological investigations using portable sensors for detection of sensitive samples

Mir,

Wani,

Akhtar

et al. 2023

Heliyon

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Genome centric engineering using ZFNs, TALENs and CRISPR-Cas9 systems for trait improvement and disease control in Animals

Cited by 21 publications

References 172 publications

Establishment of FUT8 gene knockout CHO cell line with stable expression of monoclonal antibody

Establishment of FUT8 gene knockout CHO cell line with stable expression of monoclonal antibody

Utilization of Genome Editing for Livestock Resilience in Changing Environment

Advancing biological investigations using portable sensors for detection of sensitive samples

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