Intracellular Delivery OF CRISPR/Cas9 Plasmid With Zif-8 Nanocarrier to Liver Cancer Cells
Salo, Jessica (2023)
Salo, Jessica
2023
Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi-fe20230915126893
https://urn.fi/URN:NBN:fi-fe20230915126893
Tiivistelmä
Hepatocellular carcinomas are the fifth most common cancers with poor prognosis, inefficient drug therapy, and severe side effects. C-Myc regulates up to 20 % of all human genes. C-Myc overexpression is involved in 70 % of liver cancers. Downregulating the c-Myc expression may provide a potential therapeutic alternative for future liver cancer treatments.
CRISPR/Cas9 method enables simple, stable, and inexpensive gene editing. Zif-8 biomineralization has been utilized to encapsulate CRISPR/Cas9 edited plasmid due to its stable structure, fast and easy assembling in room temperature aqueous solution, and flexibility in payload sizes since they can directly grow a nanolayer protection shell on top of the payload. Zif-8 enables pH-mediated cargo release, which can promote endosomal escape. The aims are to deliver the CRISPR/Cas9 system to knockout c-Myc genes in HCC cells with Zif-8 nanocarrier, monitor endosomal escape, analyze the downregulated c-Myc expression, and investigate the treatment effects on cancer cells.
As a result, the CRISPR/Cas9 plasmid has been successfully and efficiently delivered into HCC cells; high ratio endosomal escape after 4 hours of transfection, whereas nuclei targeting was observed at 6 hours. The transfected HCC cells with c-Myc knockout plasmid result in lower c-Myc expression than wild-type HCC cells. Results indicate the potential of intracellular c-Myc knockout with Zif-8-mediated CRISPR/Cas9 plasmid delivery in HCC cells.
Although the c-Myc expression downregulation utilization has not yet proceeded into clinical trials, it is a potential therapy for future liver cancers. The delivery method must become more accurate and suitable for further in vivo experiments.
CRISPR/Cas9 method enables simple, stable, and inexpensive gene editing. Zif-8 biomineralization has been utilized to encapsulate CRISPR/Cas9 edited plasmid due to its stable structure, fast and easy assembling in room temperature aqueous solution, and flexibility in payload sizes since they can directly grow a nanolayer protection shell on top of the payload. Zif-8 enables pH-mediated cargo release, which can promote endosomal escape. The aims are to deliver the CRISPR/Cas9 system to knockout c-Myc genes in HCC cells with Zif-8 nanocarrier, monitor endosomal escape, analyze the downregulated c-Myc expression, and investigate the treatment effects on cancer cells.
As a result, the CRISPR/Cas9 plasmid has been successfully and efficiently delivered into HCC cells; high ratio endosomal escape after 4 hours of transfection, whereas nuclei targeting was observed at 6 hours. The transfected HCC cells with c-Myc knockout plasmid result in lower c-Myc expression than wild-type HCC cells. Results indicate the potential of intracellular c-Myc knockout with Zif-8-mediated CRISPR/Cas9 plasmid delivery in HCC cells.
Although the c-Myc expression downregulation utilization has not yet proceeded into clinical trials, it is a potential therapy for future liver cancers. The delivery method must become more accurate and suitable for further in vivo experiments.
Kokoelmat
- 3111 Biolääketieteet [11]