Genome Editing in Somatic Cells Using Zinc Finger Nucleases and Transcription Activator-Like Effector Nucleases

Today we can use multiple of endonucleases for genome editing which has become very important and used in number of applications. We use sequence specific molecular scissors out of which, most important are mega nucleases, zinc finger nucleases, TALENS (Transcription Activator Like-Effector Nucleases) and CRISPR-Cas9 which is currently the most famous due to a number of reasons, they are cheap, easy to build, very specific in nature and their success rate in plants and animals is also high. Who knew that one day these CRISPR discovered as a part of immune system of bacteria will be this much worthwhile in the field of genetic engineering? This review interprets the science behind their mechanism and how several advancements were made with the passage of time to make them more efficient for the assigned job.

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Application of genome-editing systems to enhance available pig resources for agriculture and biomedicine

Reproduction Fertility and Development ◽

10.1071/rd19273 ◽

2020 ◽

Vol 32 (2) ◽

pp. 40

Author(s):

Kiho Lee ◽

Kayla Farrell ◽

Kyungjun Uh

Keyword(s):

Genetic Engineering ◽

Genome Editing ◽

Direct Injection ◽

Somatic Cells ◽

Cost Effective ◽

Genetically Engineered ◽

Zinc Finger Nucleases ◽

Transcription Activator ◽

Developmental Defects ◽

Site Specific

Traditionally, genetic engineering in the pig was a challenging task. Genetic engineering of somatic cells followed by somatic cell nuclear transfer (SCNT) could produce genetically engineered (GE) pigs carrying site-specific modifications. However, due to difficulties in engineering the genome of somatic cells and developmental defects associated with SCNT, a limited number of GE pig models were reported. Recent developments in genome-editing tools, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system, have markedly changed the effort and time required to produce GE pig models. The frequency of genetic engineering in somatic cells is now practical. In addition, SCNT is no longer essential in producing GE pigs carrying site-specific modifications, because direct injection of genome-editing systems into developing embryos introduces targeted modifications. To date, the CRISPR/Cas9 system is the most convenient, cost-effective, timely and commonly used genome-editing technology. Several applicable biomedical and agricultural pig models have been generated using the CRISPR/Cas9 system. Although the efficiency of genetic engineering has been markedly enhanced with the use of genome-editing systems, improvements are still needed to optimally use the emerging technology. Current and future advances in genome-editing strategies will have a monumental effect on pig models used in agriculture and biomedicine.

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Delivery Approaches for Therapeutic Genome Editing and Challenges

Genes ◽

10.3390/genes11101113 ◽

2020 ◽

Vol 11 (10) ◽

pp. 1113 ◽

Cited By ~ 2

Author(s):

Ilayda Ates ◽

Tanner Rathbone ◽

Callie Stuart ◽

P. Hudson Bridges ◽

Renee N. Cottle

Keyword(s):

Clinical Trials ◽

Genome Editing ◽

Zinc Finger ◽

Gene Editing ◽

Zinc Finger Nucleases ◽

Systemic Delivery ◽

Transcription Activator ◽

Therapeutic Gene ◽

Major Barrier ◽

Associated Proteins

Impressive therapeutic advances have been possible through the advent of zinc-finger nucleases and transcription activator-like effector nucleases. However, discovery of the more efficient and highly tailorable clustered regularly interspaced short palindromic repeats (CRISPR) and associated proteins (Cas9) has provided unprecedented gene-editing capabilities for treatment of various inherited and acquired diseases. Despite recent clinical trials, a major barrier for therapeutic gene editing is the absence of safe and effective methods for local and systemic delivery of gene-editing reagents. In this review, we elaborate on the challenges and provide practical considerations for improving gene editing. Specifically, we highlight issues associated with delivery of gene-editing tools into clinically relevant cells.

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Genome Editing Using Zinc-Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs)

Targeted Genome Editing Using Site-Specific Nucleases ◽

10.1007/978-4-431-55227-7_1 ◽

2014 ◽

pp. 3-24 ◽

Cited By ~ 2

Author(s):

Hiroshi Ochiai ◽

Takashi Yamamoto

Keyword(s):

Genome Editing ◽

Zinc Finger ◽

Zinc Finger Nucleases ◽

Transcription Activator

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Faculty Opinions recommendation of High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13158957.14496054 ◽

2011 ◽

Author(s):

Gregg Homanics

Keyword(s):

Genome Editing ◽

Zinc Finger ◽

High Frequency ◽

Zinc Finger Nucleases

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Faculty Opinions recommendation of Site-specific genome editing in Plasmodium falciparum using engineered zinc-finger nucleases.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717963022.793465051 ◽

2012 ◽

Author(s):

Frank Seeber

Keyword(s):

Plasmodium Falciparum ◽

Genome Editing ◽

Zinc Finger ◽

Zinc Finger Nucleases ◽

Site Specific

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Mini-Review Regarding the Applicability of Genome Editing Techniques Developed for Studying Infertility

Diagnostics ◽

10.3390/diagnostics11020246 ◽

2021 ◽

Vol 11 (2) ◽

pp. 246

Author(s):

Bogdan Doroftei ◽

Ovidiu-Dumitru Ilie ◽

Maria Puiu ◽

Alin Ciobica ◽

Ciprian Ilea

Keyword(s):

Motor Activity ◽

Experimental Model ◽

Zinc Finger ◽

Biomedical Research ◽

Zinc Finger Nucleases ◽

Loss Of Function ◽

Transcription Activator ◽

Phenotypic Changes ◽

Wide Range ◽

Short Palindromic Repeat

Infertility is a highly debated topic today. It has been long hypothesized that infertility has an idiopathic cause, but recent studies demonstrated the existence of a genetic substrate. Fortunately, the methods of editing the human genome proven to be revolutionary. Following research conducted, we identified a total of 21 relevant studies; 14 were performed on mice, 5 on zebrafish and 2 on rats. We concluded that over forty-four genes in total are dispensable for fertility in both sexes without affecting host homeostasis. However, there are genes whose loss-of-function induces moderate to severe phenotypic changes in both sexes. There were situations in which the authors reported infertility, exhibited by the experimental model, or other pathologies such as cryptorchidism, cataracts, or reduced motor activity. Overall, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 are techniques that offer a wide range of possibilities for studying infertility, even to create mutant variants. It can be concluded that ZFNs, TALENs, and CRISPR/Cas9 are crucial tools in biomedical research.

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Concepts of Molecular Plant Breeding and Genome Editing

Advances in Environmental Engineering and Green Technologies - Molecular Plant Breeding and Genome Editing Tools for Crop Improvement ◽

10.4018/978-1-7998-4312-2.ch001 ◽

2021 ◽

pp. 1-15

Keyword(s):

Plant Breeding ◽

Genome Editing ◽

Genetic Linkage ◽

Selection Process ◽

Crop Improvement ◽

Zinc Finger Nucleases ◽

Induced Mutations ◽

Transcription Activator ◽

Targeted Gene Editing ◽

Breeding Techniques

Traditional plant breeding depends on spontaneous and induced mutations available in the crop plants. Such mutations are rare and occur randomly. By contrast, molecular breeding and genome editing are advanced breeding techniques that can enhance the selection process and produce precisely targeted modifications in any crop. Identification of molecular markers, based on SSRs and SNPs, and the availability of high-throughput (HTP) genotyping platforms have accelerated the process of generating dense genetic linkage maps and thereby enhanced application of marker-assisted breeding for crop improvement. Advanced molecular biology techniques that facilitate precise, efficient, and targeted modifications at genomic loci are termed as “genome editing.” The genome editing tools include “zinc-finger nucleases (ZNFs),” “transcription activator-like effector nucleases (TALENs),” oligonucleotide-directed mutagenesis (ODM), and “clustered regularly interspersed short palindromic repeats (CRISPER/Cas) system,” which can be used for targeted gene editing. Concepts of molecular plant breeding and genome editing systems are presented in this chapter.

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