posted on 2021-11-03, 18:42authored byIman Hashemzadeh, Akbar Hasanzadeh, Fatemeh Radmanesh, Beheshteh Khodadadi Chegeni, Elaheh Sadat Hosseini, Jafar Kiani, Ali Shahbazi, Marzieh Naseri, Yousef Fatahi, Helena Nourizadeh, Behjat Kheiri Yeghaneh Azar, Amir R. Aref, Yong Liu, Michael R. Hamblin, Mahdi Karimi
Carbon
dots (CDs) have become the focus of many studies due to
their outstanding optical properties and good biocompatibility. We
investigated their potential application to produce a smart and highly
efficient yet nontoxic nanovector for gene delivery. This was achieved
by conjugating PEI1.8k-functionalized CDs (synthesized
by one-step microwave-assisted pyrolysis) with arginine-disulfide
linkers to produce CD-PEI1.8k-Arg nanoparticles. This nanovector
could deliver p-CRISPR (9.3 kb) into different types of cell lines
with higher efficiency compared to native PEI1.8k or PEI25k. CD-PEI1.8k-Arg also maintained its outstanding
transfection efficiency at a high serum concentration and low p-CRISPR
dose, compared to PEI25k, which was ineffective under those
conditions. Additionally, CD-PEI1.8k-Arg could knock out
the GFP gene with great efficiency by delivering the required components
of CRISPR/Cas9, including a plasmid encoding Cas9, sgRNA targeting
GFP, and Cas9/sgRNA ribonucleoproteins (RNPs) into the HEK 293T-GFP
cells. Moreover, the nanoparticles showed potential for the local
delivery of p-CRISPR into brain tissue. The remarkable properties
of CD-PEI1.8k-Arg could enable the development of a safe,
highly efficient gene-delivery nanovector for the treatment of various
diseases in the near future.