High‐throughput sequencing revealed low‐efficacy genome editing using Cas9 <scp>RNPs</scp> electroporation and single‐celled microinjection provided an alternative to deliver <scp>CRISPR</scp> reagents into <i>Euglena gracilis</i>

Chen, Zhen-Fan; Zhu, Jiayi; Chen, Zixi; Du, Ming; Yao, Rao; Fu, Wen; Lei, Anping; Wang, Jiangxin

doi:10.1111/pbi.13915

Cited by 7 publications

(2 citation statements)

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Section: Electroporationmentioning

confidence: 99%

“…[76] The resulting CRTP1 mutant provides a good candidate strain for studying carotenoid metabolism in E. gracilis. [68] The microinjection technique is good at directly and rapidly introducing gene fragments into single cells. However, it also requires sophisticated instruments and professional person to operate, and each experiment can only process hundreds of cells at most, so it is impractical for in vivo applications that need to edit millions of cells at the same time.…”

Section: Introductionmentioning

confidence: 99%

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Development of delivery strategies for CRISPR‐Cas9 genome editing

Liu

Yang

Xing

et al. 2023

BMEMat

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The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR‐related protein 9 (Cas9) genome editing system has attracted much attention due to its powerful genome editing capacity. However, CRISPR‐Cas9 components are easily degraded by acids, enzymes, and other substances in the body fluids after entering the organism, thus efficiently delivering the CRISPR‐Cas9 system into targeted organs or cells has been a central theme for promoting the application of CRISPR‐Cas9 technology. Although several physical methods and viral vectors have been developed for CRISPR‐Cas9 delivery, their clinical application still suffers from disadvantages, such as the risks of mutagenesis, cell damage, and poor specificity. As an alternative, non‐viral nanocarriers hold great promise for circumventing these challenges. Furthermore, with aim to realize more efficient and precise genome editing and reduce the undesirable side effects, stimuli‐responsive nanocarriers are designed for the spatiotemporal CRISPR‐Cas9 delivery in responsive to various stimuli. In this review, we will summarize the recent progress in delivery strategies for CRISPR‐Cas9 genome editing. The mechanisms and advantages of these strategies were reviewed, providing a comprehensive review of the rational design of materials and techniques for efficient and precise genome editing. At last, the potential challenges of current CRISPR‐Cas9 delivery are discussed.

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Section: Electroporationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%