Abstract:
:Fluorescent proteins have revolutionized biomedical research as they are easy to use for protein tagging, cope without fixation or permeabilization, and thus, enable live cell imaging in various models. Current methods allow easy and quick integration of fluorescent markers to endogenous genes of interest. In this review, we introduce the three central methods, zinc finger nucleases (ZFNs), transcription activator-like effectors (TALENs), and CRISPR, that have been widely used to manipulate cells or organisms. Focusing on CRISPR technology, we give an overview on homology-directed repair (HDR)-, microhomology-mediated end joining (MMEJ)-, and nonhomologous end joining (NHEJ)-based strategies for the knock-in of markers, figure out recent developments of the technique for highly efficient knock-in, and demonstrate pros and cons. We highlight the unique aspects of fluorescent protein knock-ins and pinpoint specific improvements and perspectives, like the combination of editing with stem cell derived organoid development.
journal_name
Trends Cell Bioljournal_title
Trends in cell biologyauthors
Bukhari H,Müller Tdoi
10.1016/j.tcb.2019.08.004subject
Has Abstractpub_date
2019-11-01 00:00:00pages
912-928issue
11eissn
0962-8924issn
1879-3088pii
S0962-8924(19)30140-0journal_volume
29pub_type
杂志文章,评审abstract::Microbiology has a long way to go. Microbes are ubiquitous, and all other life forms in the biosphere exist solely because of them, but, as less than 1% of microorganisms can be grown in the laboratory, more than a century of research has revealed only the tip of the iceberg concerning this most crucial of life scienc...
journal_title:Trends in cell biology
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journal_title:Trends in cell biology
pub_type: 杂志文章,评审
doi:10.1016/j.tcb.2017.07.006
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journal_title:Trends in cell biology
pub_type: 杂志文章,评审
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journal_title:Trends in cell biology
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