Fluorogenic RNA Imaging Sensors

Presenter

Qinge Liu

Campus

UMass Amherst

Sponsor

Mingxu You, Department of Chemistry, UMass Amherst

Schedule

Session 4, 2:30 PM - 3:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A66, Campus Center Auditorium, Row 4 (A61-A80) [Poster Location Map]

Abstract

Multiplexing beyond two or three RNA targets in living cells remains a technical challenge currently due to various reasons such as limited orthogonal tags and spectrum overlap. The research focuses on a new way to achieve multiplex cellular imaging through fluorogenic RNA imaging sensors. The Fluorogenic RNA(FR) aptamers exhibit the ability to selectively bind to specific non-fluorescent dyes, thereby amplifying their fluorescence. Multiple orthogonal FR/dye pairs can be used to hybridize with different cellular endogenous RNA targets, which when they bind activates the fluorescence signal of the dyes. The dye molecules are cell membrane permeable and can be easily imaged and stripped for the sequential probing of multiple RNA species in living cells. 

This project utilizes orthogonal FR/dye pairs, specifically the Pepper aptamer and HBC620 dye for the detection of Theophylline and Neomycin small molecules. The Pepper aptamer is engineered together with Theophylline aptamer and with Neomycin aptamer in creating sensors. The structure of pepper aptamer is disrupted prior to target binding ensuring a minimal fluorescence signal when it interacts with the HBC620 dye. Upon binding with the target small molecule, the sensor undergoes a conformational change, thereby activating the fluorescence signal. The Pepper/Theophylline(PepTheo) sensor and Pepper/Neomycin(PepNeo) sensor have successfully resulted in a 2-fold fluorescence enhancement upon binding with target small molecules. The methodology can also be extended to RNA detection and be potentially utilized in examining the dynamics of gene expression profiles and cellular heterogeneity on a broader scale.

Keywords

RNA, Fluorescence, Sensor

Research Area

Chemistry and Materials Science

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