TY - JOUR
T1 - Highly Photoluminescent Nitrogen- And Zinc-Doped Carbon Dots for Efficient Delivery of CRISPR/Cas9 and mRNA
AU - Hasanzadeh, Akbar
AU - Radmanesh, Fatemeh
AU - Hosseini, Elaheh Sadat
AU - Hashemzadeh, Iman
AU - Kiani, Jafar
AU - Nourizadeh, Helena
AU - Naseri, Marzieh
AU - Fatahi, Yousef
AU - Chegini, Fateme
AU - Madjd, Zahra
AU - Beyzavi, Ali
AU - Kowalski, Piotr S.
AU - Karimi, Mahdi
N1 - Publisher Copyright:
©
PY - 2021/8/18
Y1 - 2021/8/18
N2 - Safe and efficient delivery of CRISPR/Cas9 systems is still a challenge. Here we report the development of fluorescent nitrogen- and zinc-doped carbon dots (N-Zn-doped CDs) using one-step microwave-aided pyrolysis based on citric acid, branched PEI25k, and different zinc salts. These versatile nanovectors with a quantum yield of around 60% could not only transfect large CRISPR plasmids (∼9 kb) with higher efficiency (80%) compared to PEI25k and lipofectamine 2000 (Lipo 2K), but they also delivered mRNA into HEK 293T cells with the efficiency 20 times greater than and equal to that of PEI25k and Lipo 2K, respectively. Unlike PEI25k, N-Zn-doped CDs exhibited good transfection efficiency even at low plasmid doses and in the presence of 10% fetal bovine serum (FBS). Moreover, these nanovectors demonstrated excellent efficiency in GFP gene disruption by transferring plasmid encoding Cas9 and sgRNA targeting GFP as well as Cas9/sgRNA ribonucleoproteins into HEK 293T-GFP cells. Hence, N-Zn-doped CDs with remarkable photoluminescence properties and high transfection efficiency in the delivery of both CRISPR complexes and mRNA provide a promising platform for developing safe, efficient, and traceable delivery systems for biological research.
AB - Safe and efficient delivery of CRISPR/Cas9 systems is still a challenge. Here we report the development of fluorescent nitrogen- and zinc-doped carbon dots (N-Zn-doped CDs) using one-step microwave-aided pyrolysis based on citric acid, branched PEI25k, and different zinc salts. These versatile nanovectors with a quantum yield of around 60% could not only transfect large CRISPR plasmids (∼9 kb) with higher efficiency (80%) compared to PEI25k and lipofectamine 2000 (Lipo 2K), but they also delivered mRNA into HEK 293T cells with the efficiency 20 times greater than and equal to that of PEI25k and Lipo 2K, respectively. Unlike PEI25k, N-Zn-doped CDs exhibited good transfection efficiency even at low plasmid doses and in the presence of 10% fetal bovine serum (FBS). Moreover, these nanovectors demonstrated excellent efficiency in GFP gene disruption by transferring plasmid encoding Cas9 and sgRNA targeting GFP as well as Cas9/sgRNA ribonucleoproteins into HEK 293T-GFP cells. Hence, N-Zn-doped CDs with remarkable photoluminescence properties and high transfection efficiency in the delivery of both CRISPR complexes and mRNA provide a promising platform for developing safe, efficient, and traceable delivery systems for biological research.
UR - https://www.scopus.com/pages/publications/85111302748
U2 - 10.1021/acs.bioconjchem.1c00309
DO - 10.1021/acs.bioconjchem.1c00309
M3 - Article
C2 - 34278778
AN - SCOPUS:85111302748
SN - 1043-1802
VL - 32
SP - 1875
EP - 1887
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 8
ER -