Poly-sgRNA/siRNA ribonucleoprotein nanoparticles for targeted gene disruption.
Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9) can be used for the specific disruption of a target gene to permanently suppress the expression of the protein encoded by the target gene. Efficient delivery of the system to an intracellular target site should be achieved to utilize the tremendous potential of the genome-editing tool in biomedical applications such as the knock-out of disease-related genes and the correction of defect genes. Here, we devise polymeric CRISPR/Cas9 system based on poly-ribonucleoprotein (RNP) nanoparticles consisting of polymeric sgRNA, siRNA, and Cas9 endonuclease in order to improve the delivery efficiency. When delivered by cationic lipids, the RNP nanoparticles built with chimeric poly-sgRNA/siRNA sequences generate multiple sgRNA-Cas9 RNP complexes upon the Dicer-mediated digestion of the siRNA parts, leading to more efficient disruption of the target gene in cells and animal models, compared with the monomeric sgRNA-Cas9 RNP complex.
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
Jong Seong
JS
Ha
Department of Chemical Engineering, University of Seoul, Seoul 02504, Republic of Korea.
Jae Sung
JS
Lee
Department of Chemical Engineering, University of Seoul, Seoul 02504, Republic of Korea.
Jaepil
J
Jeong
Department of Chemical Engineering, University of Seoul, Seoul 02504, Republic of Korea.
Hejin
H
Kim
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
Juyoung
J
Byun
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; School of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea.
Sang Ah
SA
Kim
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
Hee Jae
HJ
Lee
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, KIST School, Korea University of Science and Technology (UST), Seoul 02792, Republic of Korea.
Hak Suk
HS
Chung
Department of Chemical Engineering, University of Seoul, Seoul 02504, Republic of Korea. Electronic address: jblee@uos.ac.kr.
Jong Bum
JB
Lee
The Center for Theragnosis, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, KIST School, Korea University of Science and Technology (UST), Seoul 02792, Republic of Korea. Electronic address: drahn@kist.re.kr.
Dae-Ro
DR
Ahn
D000818:Animals; D064113:CRISPR-Cas Systems; D002470:Cell Survival / Q000187:drug effects; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004337:Drug Carriers; D004720:Endonucleases / Q000737:chemistry*; D018390:Gene Targeting; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D053758:Nanoparticles / Q000737:chemistry*; D009368:Neoplasm Transplantation; D010316:Particle Size; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry*; D043244:Ribonuclease III / Q000737:chemistry; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000633:toxicity
D004337:Drug Carriers; D000094704:RNA, Guide, CRISPR-Cas Systems; D034741:RNA, Small Interfering; D012261:Ribonucleoproteins; D004720:Endonucleases; D043244:Ribonuclease III
CRISPR/Cas9; Gene delivery; Gene disruption; Poly-ribonucleoprotein; Rolling circle amplification