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Highly accurate small RNA sequencing of single cells RealSeq SC

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R43HG009863-01
Agency Tracking Number: R43HG009863
Amount: $281,170.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: 172
Solicitation Number: PA16-302
Solicitation Year: 2016
Award Year: 2017
Award Start Date (Proposal Award Date): 2017-07-10
Award End Date (Contract End Date): 2019-04-30
Small Business Information
Santa Cruz, CA 95060-5790
United States
DUNS: 013494781
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 (831) 426-7700
Business Contact
Phone: (831) 426-7700
Research Institution

The goal of this grant application is to develop the first commercially available library preparation kit for profiling
small RNAs from single cells using NGS methods Single cell analyses of mRNA have allowed the
identification of crucial differences between cells that were otherwise considered identical These findings have
shown that there is intrinsic noise in the regulation of gene expression that plays an important role in
determining cell fates Unfortunately there is currently a lack of information about the cell to cell variability of
levels of small RNAs including microRNAs Indeed there is no commercially available library preparation kit
for small RNAs that can profile single cells We propose to further develop our library preparation technology
RealSeq AC to be able to quantify small RNAs from single cells RealSeq AC uses a scheme involving a
single combo adapter and circularization that accurately quantifies over of all miRNAs detected
compared to from the best competitor kit To adapt this technology for single cell sequencing we will test
three separate strategies to dramatically reduce the presence of adapter dimers in the library as well as
possible use of miRNA pre amplification to reduce the number of PCR cycles needed We will develop a
protocol that performs all steps from cell lysis to final purification of amplified libraries in a single tube This
technology will allow the accurate quantification of small RNAs from single cells Health relatedness narrative
Transcriptomic analysis of single cells has shown that there is a high cell to cell variability in gene expression
among apparently identical cells in a population However a similar analysis is not currently possible for levels
of small RNAs which play key roles in regulating gene expression The goal of this grant application is to
develop the first commercially available library preparation kit for profiling microRNAs from single cells using
modern high throughput sequencing methods Understanding how individual cells vary in their small RNA
profiles is of central importance to understanding cancer progression and the evolution of malignant tumor

* Information listed above is at the time of submission. *

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