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564 学报 Journal of China Pharmaceutical University 2025, 56(5): 557 − 565 第 56 卷

白可能引起免疫反应，通过优化载体粒径和表面性 ated proximity ligation assay[J]. J Am Chem Soc, 2018,
质可降低清除率 ，从而延长体内循环时间;（3）靶 140(36): 11293-11301.
[55]
[13] Ma HH, Tu LC, Naseri A, et al. Multiplexed labeling of genom-
向特定组织的难度，需在载体上引入特异性适体以 ic loci with dCas9 and engineered sgRNAs using CRISPRain-
提高靶向性 。相比之下，固定组织中的成像难度 bow[J]. Nat Biotechnol, 2016, 34(5): 528-530.
[56]
较低，但目前相关研究和应用较少。将 CRISPR/Cas9 [14] Wang HF, Nakamura M, Abbott TR, et al. CRISPR-mediated
live imaging of genome editing and transcription[J]. Science,
成像系统引入固定组织中，有望作为 FISH 方法的 2019, 365(6459): 1301-1305.
补充，进一步拓展临床诊断的应用范围。 [15] Saka SK, Wang Y, Kishi JY, et al. Immuno-SABER enables
未来，CRISPR/Cas9 原位成像技术将围绕优化 highly multiplexed and amplified protein imaging in tissues[J].
Nat Biotechnol, 2019, 37(9): 1080-1090.
sgRNA 结构、探索高效递送策略、开发优质荧光染
[16] Chen BH, Gilbert LA, Cimini BA, et al. Dynamic imaging of
料、设计新型信号放大策略及推动临床应用展开。 genomic loci in living human cells by an optimized
CRISPR/Cas9 原位成像技术在生物成像领域具有 CRISPR/Cas system[J]. Cell, 2013, 155(7): 1479-1491.
[17] Ma HH, Naseri A, Reyes-Gutierrez P, et al. Multicolor CRISPR
广阔的应用前景，随着技术的不断发展和完善，其
labeling of chromosomal loci in human cells[J]. Proc Natl Acad
将为生物学研究和临床诊断带来更多可能性。 Sci U S A, 2015, 112(10): 3002-3007.
[18] Chen BH, Hu J, Almeida R, et al. Expanding the CRISPR imag-
ing toolset with Staphylococcus aureus Cas9 for simultaneous
References
imaging of multiple genomic loci[J]. Nucleic Acids Res, 2016,
[1] Eng CL, Lawson M, Zhu Q, et al. Transcriptome-scale super-re- 44(8): e75.
solved imaging in tissues by RNA seqFISH[J]. Nature, 2019, [19] Chen BH, Zou W, Xu HY, et al. Efficient labeling and imaging
568(7751): 235-239. of protein-coding genes in living cells using CRISPR-Tag[J].
[2] Wu XF, Wang R, Qi SJ, et al. Rational design of a highly selec- Nat Commun, 2018, 9(1): 5065.
tive near-infrared two-photon fluorogenic probe for imaging or- [20] Zhao Y, Cao ST, Wang Y, et al. A temporally resolved DNA
thotopic hepatocellular carcinoma chemotherapy[J]. Angew framework state machine in living cells[J]. Nat Mach Intell,
Chem Int Ed, 2021, 60(28): 15418-15425. 2023, 5(9): 980-990.
[3] Kim J, Lee S, Kim Y, et al. In situ self-assembly for cancer [21] Takata H, Masuda Y, Ohmido N. CRISPR imaging reveals
therapy and imaging[J]. Nat Rev Mater, 2023, 8(11): 710-725. chromatin fluctuation at the centromere region related to cellu-
[4] Fang RX, Xia CL, Close JL, et al. Conservation and divergence lar senescence[J]. Sci Rep, 2023, 13(1): 14609.
of cortical cell organization in human and mouse revealed by [22] Takei Y, Shah S, Harvey S, et al. Multiplexed dynamic imag-
MERFISH[J]. Science, 2022, 377(6601): 56-62. ing of genomic loci by combined CRISPR imaging and DNA
[5] Zhang BW, Tian TR, Xiao DX, et al. Facilitating in situ tumor sequential FISH[J]. Biophys J, 2017, 112(9): 1773-1776.
imaging with a tetrahedral DNA framework-enhanced hy- [23] Guan J, Liu H, Shi XY, et al. Tracking multiple genomic ele-
bridization chain reaction probe[J]. Adv Funct Mater, 2022, ments using correlative CRISPR imaging and sequential DNA
32(16): 2109728. FISH[J]. Biophys J, 2017, 112(6): 1077-1084.
[6] Safieddine A, Coleno E, Lionneton F, et al. HT-smFISH: a cost- [24] Tanenbaum ME, Gilbert LA, Qi LS, et al. A protein-tagging
effective and flexible workflow for high-throughput single- system for signal amplification in gene expression and fluores-
molecule RNA imaging[J]. Nat Protoc, 2022, 18(1): 157-187. cence imaging[J]. Cell, 2014, 159(3): 635-646.
[7] Veselinyová D, Mašlanková J, Kalinová K, et al. Selected in [25] Ye HY, Rong ZL, Lin Y. Live cell imaging of genomic loci us-
situ hybridization methods: principles and application[J]. ing dCas9-SunTag system and a bright fluorescent protein[J].
Molecules, 2021, 26(13): 3874. Protein Cell, 2017, 8(11): 853-855.
[8] Parsons AM, Byrne S, Kooistra J, et al. G-quadruplex stabiliza- [26] Knight SC, Xie LQ, Deng WL, et al. Dynamics of CRISPR-
tion provokes DNA breaks in human PKD1, revealing a second Cas9 genome interrogation in living cells[J]. Science, 2015,
hit mechanism for ADPKD[J]. Nat Commun, 2025, 16(1): 121. 350(6262): 823-826.
[9] Niu YY, Shen B, Cui YQ, et al. Generation of gene-modified [27] Deng WL, Shi XH, Tjian R, et al. CASFISH: CRISPR/Cas9-
Cynomolgus monkey via Cas9/RNA-mediated gene targeting in mediated in situ labeling of genomic loci in fixed cells[J]. Proc
one-cell embryos[J]. Cell, 2014, 156(4): 836-843. Natl Acad Sci U S A, 2015, 112(38): 11870-11875.
[10] Carroll D. Genome engineering with targetable nucleases[J]. [28] Neguembor MV, Sebastian-Perez R, Aulicino F, et al.
Annu Rev Biochem, 2014, 83: 409-439. (Po)STAC (Polycistronic SunTAg modified CRISPR) enables
[11] Jinek M, Chylinski K, Fonfara I, et al. A programmable dual- live-cell and fixed-cell super-resolution imaging of multiple
RNA-guided DNA endonuclease in adaptive bacterial immuni- genes[J]. Nucleic Acids Res, 2018, 46(5): e30.
ty[J]. Science, 2012, 337(6096): 816-821. [29] Hou Y, Wang D, Lu S, et al. Optogenetic control of back-
[12] Zhang KX, Deng RJ, Teng XC, et al. Direct visualization of sin- ground fluorescence reduction for CRISPR-based genome
gle-nucleotide variation in mtDNA using a CRISPR/Cas9-medi- imaging[J]. Anal Chem, 2022, 94(24): 8724-8731.