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| Systematic Investigation of the Effects of Multiple SV40 Nuclear Localization Signal Fusion on the Genome Editing Activity of Purified SpCas9 | |
| 2022-02 | |
| Source Publication | BIOENGINEERING-BASEL
 |
| EISSN | 2306-5354 |
| Volume | 9Issue:2 |
| Status | 已发表 |
| DOI | 10.3390/bioengineering9020083 |
| Abstract | The emergence of CRISPR-Cas9 technology has revolutionized both basic and translational biomedical research. For Cas9 nuclease to exert genome editing activity, nuclear localization signal (NLS) derived from simian virus 40 (SV40) T antigen is commonly installed as genetic fusion to direct the intracellular Cas9 proteins to the nucleus of cells. Notably, previous studies have shown that multiple SV40 NLS fusion can improve the targeting activity of Cas9-derived genome-editing and base-editing tools. In addition, the multi-NLS fusion can increase the intracellular activity of Cas9 in the forms of both constitutive expression and directly delivered Cas9-guide RNA ribonucleoprotein (RNP) complex. However, the relationship between NLS fusion and intracellular Cas9 activity has not been fully understood, including the dependency of activity on the number or organization of NLS fusion. In the present study, we constructed and purified a set of Streptococcus pyogenes Cas9 (SpCas9) variants containing one to four NLS repeats at the N- or C-terminus of the proteins and systematically analyzed the effects of multi-NLS fusion on the activity of SpCas9 RNPs. It was found that multi-NLS fusion could improve the intracellular activity as lipofected or nucleofected Cas9 RNPs. Importantly, multi-NLS fusion could enhance the genome-editing activity of SpCas9 RNPs in primary and stem/progenitor cells and mouse embryos. |
| Keyword | CRISPR-Cas9 Cas9 ribonucleoproteins (RNPs) nuclear localization signal (NLS) genome editing |
| URL | 查看原文 |
| Indexed By | SCI ; SCIE |
| Language | 英语 |
| WOS Research Area | Biotechnology & Applied Microbiology ; Engineering |
| WOS Subject | Biotechnology & Applied Microbiology ; Engineering, Biomedical |
| WOS ID | WOS:000771659000001 |
| Publisher | MDPI |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/165001 |
| Collection | 生命科学与技术学院_硕士生 免疫化学研究所_PI研究组_刘佳组 |
| Corresponding Author | Liu, Jia |
| Affiliation | 1.Shanghai Tech Univ, Shanghai Inst Adv Immunochem Studies, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 2.Lab Prot Design & Immunoengn LPDI STI EPFL, CH-1015 Lausanne, Switzerland 3.Swiss Inst Bioinformat SIB, CH-1015 Lausanne, Switzerland |
| First Author Affilication | Shanghai Institute for Advanced Immunochemical Studies |
| Corresponding Author Affilication | Shanghai Institute for Advanced Immunochemical Studies |
| First Signature Affilication | Shanghai Institute for Advanced Immunochemical Studies |
| Recommended Citation GB/T 7714 | Shui, Sailan,Wang, Shaojie,Liu, Jia. Systematic Investigation of the Effects of Multiple SV40 Nuclear Localization Signal Fusion on the Genome Editing Activity of Purified SpCas9[J]. BIOENGINEERING-BASEL,2022,9(2). |
| APA | Shui, Sailan,Wang, Shaojie,&Liu, Jia.(2022).Systematic Investigation of the Effects of Multiple SV40 Nuclear Localization Signal Fusion on the Genome Editing Activity of Purified SpCas9.BIOENGINEERING-BASEL,9(2). |
| MLA | Shui, Sailan,et al."Systematic Investigation of the Effects of Multiple SV40 Nuclear Localization Signal Fusion on the Genome Editing Activity of Purified SpCas9".BIOENGINEERING-BASEL 9.2(2022). |
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