Author: Long T. Nguyen; Brianna M Smith; Piyush K. Jain
Title: Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection Document date: 2020_4_14
ID: n5kpvsvg_13
Snippet: We optimized the previously developed CRISPR-based detection assays 1,5,6 and combined them with our engineered crRNA+3'DNA7 to create a CRISPR-ENHANCE (Enhanced analysis of nucleic acids with crRNA extensions) technology or referred here as ENHANCE. To validate the ENHANCE technology, we first selected a clinically relevant nucleic acid biomarker, Prostate Cancer Antigen 3 (PCA3/DD3), which is one of the most overexpressed genes in prostate canc.....
Document: We optimized the previously developed CRISPR-based detection assays 1,5,6 and combined them with our engineered crRNA+3'DNA7 to create a CRISPR-ENHANCE (Enhanced analysis of nucleic acids with crRNA extensions) technology or referred here as ENHANCE. To validate the ENHANCE technology, we first selected a clinically relevant nucleic acid biomarker, Prostate Cancer Antigen 3 (PCA3/DD3), which is one of the most overexpressed genes in prostate cancer tissue and excreted in patients' urine. Consequently, elevated PCA3 levels during prostate cancer progression has become a widely targeted biomarker for detection. [21] [22] [23] [24] To determine the limit of detection of PCA3 using our ENHANCE technology, we spiked the PCA3 cDNA into synthetic urine and investigated how this clinically-relevant environment affects the activity of Cas12a.
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