ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering
Open accessAbstract
Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) comprise a powerful class of tools that are redefining the boundaries of biological research. These chimeric nucleases are composed of programmable, sequence-specific DNA-binding modules linked to a nonspecific DNA cleavage domain. ZFNs and TALENs enable a broad range of genetic modifications by inducing DNA double-strand breaks that stimulate error-prone nonhomologous end joining or homology-directed repair at specific genomic locations. Here, we review achievements made possible by site-specific nuclease technologies and discuss applications of these reagents for genetic analysis and manipulation. In addition, we highlight the therapeutic potential of ZFNs and TALENs and discuss future prospects for the field, including the emergence of clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas-based RNA-guided DNA endonucleases.
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The CRISPR/Cas9 system has an advantage in size compared with TALENs.
“Fortunately, the CRISPR/Cas9 system has the advantage in size compared with TALENs 22 .”
From Harnessing the CRISPR/Cas9 system to disrupt latent HIV-1 provirus (2013) · cites this paper as [22]
The Future directions section says the large size of TALENs may limit delivery by size-restricted vectors. It does not compare TALEN size with CRISPR/Cas9 size, so it supports the TALEN size concern but not the claimed comparative advantage. (LLM, conf=0.82)
Experimental — accuracy not yet measuredThese nucleases induce DNA double-strand breaks at precise locations in the host genome, facilitating integration of the cassette by non-homologous end-joining (NHEJ) or homology-directed repair (HDR).
“These nucleases induce DNA double-strand breaks at precise locations into the host genome, facilitating the integration of the cassette by non-homologous end-joining (NHEJ) or homology-directed repair (HDR) (Gaj et al., 2013).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Gaj et al., 2013
TALEN technique has already been widely used for genome editing in a variety of cells, plants, model organisms, livestock and even human cells.
“In a short time after discovery, this technique has already been widely used for genome editing in a variety of cells, plants, model organisms, livestock and even human cells [66,67].”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [67]
The discovery that TALEs use a simple modular code for DNA recognition has provided an alternative platform for genome editing and gene disruption.
“The discovery that TALEs use a simple modular code for DNA recognition has provided an alternative platform for genome editing and gene disruption.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [67]
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“Alternatively, programmable nucleases, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), allow more precise gene editing in model organisms, particularly in mammalian and human systems (Gaj et al., 2013; Kim and Kim, 2014).”
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- Authors
- Gaj, T., Gersbach, C. A., & Barbas, C. F.
- Affiliations
- Scripps Research Institute (US)
- Duke University (US)
- Corresponding author
- Barbas, Carlos F.
- Venue
- Trends in Biotechnology
- Publisher
- Elsevier BV
- Published
- 2013-07
- Volume
- 31
- Issue
- 7
- Pages
- 397-405
- ISSN
- 0167-7799 (ISSN-L)
- PMC
- PMC3694601
- OpenAlex
- W2112444085
- Cited by
- 3,820
- References
- 119
- Citations by year
- 2026: 142 · 2025: 222 · 2024: 254 · 2023: 266 · 2022: 314 · 2021: 302 · 2020: 266 · 2019: 303 · 2018: 287 · 2017: 300 · 2016: 320 · 2015: 373 · 2014: 352 · 2013: 114 · 2012: 1
- FWCI
- 35.56
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Trends in biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (1dp2od008586-01)
- National Cancer Institute (DP1CA174426)
- National Institute of General Medical Sciences (T32GM080209)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- zinc finger nucleases, TALEN, CRISPR-Cas, genome editing, site-specific DNA cleavage, DNA double-strand breaks, homology-directed repair, non-homologous end joining, gene targeting, gene therapy, RNA-guided DNA endonuclease
- MeSH terms
- Biological Therapy, Deoxyribonucleases, Genetic Engineering, Molecular Biology, Recombinant Proteins, Zinc Fingers, Clustered Regularly Interspaced Short Palindromic Repeats
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