Efficient targeted insertion of large DNA fragments without DNA donors

Wang, Jinlin; Zhang, He; Wang, Guoquan; Zhang, Ruiwen; Duan, Junyi; Gao, Pan; Lei, Xinlin; Qiu, Houyuan; Chuan-ping, Zhang; Zhang, Ying; Yin, Hao

doi:10.1038/s41592-022-01399-1

Cited by 80 publications

(55 citation statements)

References 48 publications

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“…Our ultimate goal was to develop tools for endogenous mRNA imaging in live animals. It has been previously reported that the knock-in of short sequences into the genome is far more efficient than those of longer sequences 18 . The MASS exploits this advantage as only 8xMS2 (350 nt) needs to be inserted into a genomic locus, thus overcoming the previous obstacle of the requirement of inserting a long 1,300 nt 24xMS2 into the genome for live-cell imaging of endogenous mRNA.…”

Section: Resultsmentioning

confidence: 99%

Enhanced Single RNA Imaging Reveals Dynamic Gene Expression in Live Animals

Gao

et al. 2022

Preprint

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Imaging endogenous mRNAs in live animals is technically challenging. Here we describe an MS2 based signal Amplification with Suntag System that enables live-cell RNA imaging of high temporal resolution and with 8xMS2 stem-loops, which overcomes the obstacle of inserting a 1,300 nt 24xMS2 into the genome for the imaging of endogenous mRNAs. Using this tool we were able to image the activation of gene expression and the dynamics of endogenous mRNAs in the epidermis of live C. elegans.

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Section: Resultsmentioning

confidence: 99%

Enhanced Single RNA Imaging Reveals Dynamic Gene Expression in Live Animals

Gao

et al. 2022

Preprint

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“…I-CreI plays a critical role in the localization and occupancy of the catalytic metal ions, which is crucial for the DNA cleavage ( Prieto et al, 2018 ). Wang et al (2022) developed a transgenic Xenopus tropicalis line which is used for evaluating the potential effects of I-SceI mediated transgenesis and further understanding its mechanisms. The fusion of transcription activator-like effector (TALE) DNA-binding domains to MNs dramatically increases the efficiency by 35-fold compared to standalone MNs to modify T-cells receptor alpha ( Boissel et al, 2014a ).…”

Section: Nuclease-based Genome Engineering Technologiesmentioning

confidence: 99%

“…Recent efforts stepwise improved the efficiency of PE system to PEmax system ( Chen et al, 2021 ) and engineer pegRNA known as epegRNA ( Nelson et al, 2022b ). PE makes possible the accurate insertion up to 1 kb ( Wang et al, 2022a ) and the deletion of up to 10 kb ( Choi et al, 2022 ) DNA fragment.…”

Section: Nickase-based Genome Engineering Technologiesmentioning

confidence: 99%

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Mbakam²,

Song³

et al. 2022

Front. Genome Ed.

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Advancements in genome editing make possible to exploit the functions of enzymes for efficient DNA modifications with tremendous potential to treat human genetic diseases. Several nuclease genome editing strategies including Meganucleases (MNs), Zinc Finger Nucleases (ZFNs), Transcription Activator-like Effector Nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated proteins (CRISPR-Cas) have been developed for the correction of genetic mutations. CRISPR-Cas has further been engineered to create nickase genome editing tools including Base editors and Prime editors with much precision and efficacy. In this review, we summarized recent improvements in nuclease and nickase genome editing approaches for the treatment of genetic diseases. We also highlighted some limitations for the translation of these approaches into clinical applications.

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“…Subsequently, the percentages for each category of variants within all different variants are marked on the pie graph. It is also clear from the accumulated knowledge regarding these genetic diseases, certain gene defects (or given alleles) may drive a spectrum of phenotypes, whereas similar phenotypes may also be attributed to defects in different genes (1,3). Additionally, some of the defects would exhibit incomplete penetrance.…”

Section: Figurementioning

confidence: 99%

“…Traditionally known as "primary immunodeficiencies", these defects are now commonly designated as inborn errors of immunity (IEI), a term better reflecting the presentation of a broad spectrum of immune-related phenotypes. Indeed, IEI patients can feature not only susceptibility to infections, but also autoimmunity/autoinflammation, allergy and malignancy (1)(2)(3). Additionally, even patients with the same mutations may manifest heterogeneous phenotypes.…”

Section: Introductionmentioning

confidence: 99%

Precise somatic genome editing for treatment of inborn errors of immunity

2022

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Rapid advances in high throughput sequencing have substantially expedited the identification and diagnosis of inborn errors of immunity (IEI). Correction of faulty genes in the hematopoietic stem cells can potentially provide cures for the majority of these monogenic immune disorders. Given the clinical efficacies of vector-based gene therapies already established for certain groups of IEI, the recently emerged genome editing technologies promise to bring safer and more versatile treatment options. Here, we review the latest development in genome editing technologies, focusing on the state-of-the-art tools with improved precision and safety profiles. We subsequently summarize the recent preclinical applications of genome editing tools in IEI models, and discuss the major challenges and future perspectives of such treatment modalities. Continued explorations of precise genome editing for IEI treatment shall move us closer toward curing these unfortunate rare diseases.

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Efficient targeted insertion of large DNA fragments without DNA donors

Cited by 80 publications

References 48 publications

Enhanced Single RNA Imaging Reveals Dynamic Gene Expression in Live Animals

Enhanced Single RNA Imaging Reveals Dynamic Gene Expression in Live Animals

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Precise somatic genome editing for treatment of inborn errors of immunity

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