Unmet Need / Invention Novelty: Single cell RNA transcriptomics has become an important technology in research and diagnostics but can be improved by incorporating genomic readouts from the same cells whose transcriptomes are being analyzed. Sequencing both genomic DNA and messenger RNA from a single cell is crucial for associating a cell’s genotype and transcriptome, introducing opportunity in a range of applications from research and streamlined CRISPR genome editing to diagnostics. There is an unmet need to develop a high-throughput method for the efficient and simultaneous analysis of DNA and RNA from the same cell.
Technical Details: Researchers at Johns Hopkins have developed a high-throughput method to simultaneously analyze targeted DNA and whole transcriptomes from single cells. The method allows for genotyping of single cells while their transcriptome is captured in parallel. Briefly, a single cell is lysed in a droplet, in-droplet digital PCR is performed to amplify a genomic region of interest, and this amplified DNA, in addition to the mRNA, is captured by a barcoded bead for sequencing. Proof of concept studies have demonstrated successful analysis of a heterogenous population of human induced pluripotent stem cells after CRISPR/Cas9 genome editing, obviating the need for clonal selection, expansion, genotyping, and batch effect elimination normally required post CRISPR/Cas9 genome editing.
Value Proposition:
- High throughout, dual analysis of DNA and RNA from a single cell
- Efficient in-droplet PCR from cell lysate
- Applicable in non-droplet cell encapsulation methods such as FACS, gravity and microfluidics
- Advantageous to multiple applications including research, cellular barcoding, streamlined CRISPR genome editing and clinical diagnostics
Looking for Partners to: Develop & commercialize as a novel method for dual single cell analysis
Stage of Development: Pre-Clinical
Data Availability: in vitro
Inventors: Donald Zack, Claire Bell, Tza-Huei Wang, Aniruddha Kaushik, and Pengfei Zhang
Patent Status: Pending PCT application
Publication(s)/Related Technology: N/A
