Functional classification of prostate cancer‑associated miRNAs through CRISPR/Cas9‑mediated gene knockout

Abstract

The aim of the present study was to use the clustered regularly interspaced short palindromic repeats (criSPr) and criSPr-associated (cas) 9-mediated gene knockout technology for the rapid classification of the differential function of micro (mi)rnas screened using mirna expression profiling by microarray. The rational design of single guide rnas for the criSPr/Cas9 system was verified to function in human LNCaP cells with rapid and efficient target gene editing. mirna (mir)-205, mir-221, mir-222, mir-30c, mir-224, mir-455-3p, mir-23b and mir-505 were downregulated in patients with prostate cancer (Pca) and were experimentally validated to function as tumor suppressors in prostate cancer cells, affecting tumor proliferation, invasion and aerobic glycolysis. in addition, the data of the present study suggested that miR-663a and mfiR-1225-5p were upregulated in prostate cancer tissues and cell proliferation of mir-663a and miR-1225-5p knockout PCa cells was significantly lower compared with mir-nc cells. Furthermore, knockout of miR-1225-5p and miR-663a significantly decreased the lactate production in lncaP cells in vitro. in conclusion, the present study offered a simple and efficient method for rapidly classifying mirna function by applying criSPr/cas9 in lncaP cells. The present study suggested, for the first time to the best of the authors' knowledge, that the aberrant expression of mir-663a and mir-1225-5p may be involved with the progression of prostate cancer, implying their potential as candidate markers for this type of cancer. However, the precise role of mir-663a and mir-1225-5p in accelerating the development of prostate cancer and promoting tumor progression remains to be elucidated.

Lung development, repair and cancer: A study on the role of MMP20 gene in adenocarcinoma

“…Since the discovery of CRISPR-Cas9 in bacterial immune system [ 28 ], CRISPR-Cas9 has been adapted into a powerful tool for cancer genomic research [ 29 , 30 ]. Over the past decade, the CRISPR-Cas9 technology has become a popular technique for genome editing because it is rapid, cost-effective, precise and relatively easy to perform [ 31 ].…”

CRISPR/Cas: a Nobel Prize award-winning precise genome editing technology for gene therapy and crop improvement

“…The edited cell lines revealed that the miR21 knock-out significantly inhibited cancer cell proliferation, migration, and invasion, highlighting a prime target for cancer gene therapies ( Huo et al, 2017 ). To add to this, again using CRISPR/Cas9, Jiang et al (2020) were able to knock out several miRNA genes, including miR30c, miR205, and miR663a, in human LNCaP cell lines, and were able to determine that miR-1225-5p and miR-663a knock-outs decreased lactate production in LNCaP cells. However, although miRNA-related research is a quickly developing field with thousands of miRNAs already identified, most research has been focused on humans, and there is still much to learn.…”

Zastosowanie technologii CRISPR/Cas9 w leczeniu nowotworów

“…Dzięki metodzie CRISPR możliwa jest klasyfikacja mikroRNA związanych z nowotworami. W badaniu przeprowadzonym z wykorzystaniem ludzkiej linii komórkowej raka prostaty dowiedziono, że zaburzona ekspresja dwóch miRNA (miR-663a i miR-1225-5p) wpływa na progresję raka prostaty i może być wykorzystana jako potencjalny marker tego nowotworu (28). Nabranie przez komórki nowotworowe cech komórek macierzystych jest jak dotąd słabo poznane, ale wiadomo, że wpływa to na agresywny fenotyp i oporność na leczenie.…”