Abstract: Recently, advances in next-generation sequencing have arisen from the spatial isolation of each molecule into a small volume, enabling many single-molecule sequencing reactions to run in parallel. The fundamental limit to throughput with this technique is the need to isolate individual molecules on a spatial scale, so that sequencing signals are not mixed. Here we disrupt this limit, by observing that, in many cases, it is possible to accurately sequence complex mixtures of DNA and RNA species by exploiting the toolkit of modern compressed sensing and incorporating additional relational information about the relationship between many sequencing problems. This approach thus provides a dramatic increase in the density of DNA molecules in the sequencing reaction for both in-vitro and in-situ techniques.

Abstract: The present invention provides a method of identifying mRNA transcripts in the transcriptome of a cell comprising i) delivering into the cell a donor expression vector comprising nucleotides in a sequence encoding a trans-splicing barcode cassette, ii) exposing the cell to conditions such that the cell produces multiple copies of the trans-splicing barcode cassette encoded by the donor expression vector, which multiple copies of the trans-splicing barcode cassette each splice the barcode polynucleotide onto a mRNA transcript of the cell, thereby forming multiple mRNA transcripts of the cell, each spliced to the barcode polynucleotide; and iii) identifying the multiple mRNA transcripts that are spliced to the barcode polynucleotides, thereby identifying mRNA transcripts in the transcriptome of the cell.

Abstract: The present invention provides a method of identifying mRNA transcripts in the transcriptome of a cell comprising i) delivering into the cell a donor expression vector comprising nucleotides in a sequence encoding a trans-splicing barcode cassette, wherein the trans-splicing barcode cassette comprises a) a first portion, the nucleotide sequence of which encodes an intron comprising as part of its 3? end, or followed at its 3? end by a splice-site nucleotide sequence; followed at its 3? end by, b) a second portion, the nucleotide sequence of which encodes a barcode polynucleotide; followed at its 3? end by c) a third portion, which encodes a nucleotide identification element sequence, ii) exposing the cell to conditions such that the cell produces multiple copies of the trans-splicing barcode cassette encoded by the donor expression vector, which multiple copies of the trans-splicing barcode cassette each splice the barcode polynucleotide onto a mRNA transcript of the cell, thereby forming multiple mRNA transc

Abstract: The present invention provides a trans-splicing ribozyme comprising i) a targeting nucleotide sequence that is complementary to a target nucleotide sequence within a mRNA that is expressed in a cell; contiguous with ii) a catalytic RNA sequence; contiguous with iii) a donor transcript, which donor transcript comprises at least a nucleotide sequence that encodes a trans-activator, wherein when the trans-splicing ribozyme is expressed in a cell, the catalytic RNA sequence cleaves the mRNA and ligates the donor transcript to the mRNA to generate a spliced mRNA which comprises the donor transcript, such that the donor transcript is translated as part of the spliced mRNA in the cell, as well as methods of using the trans-splicing ribozyme. The present invention also provides variants of Cre and other recombinases, as well as method of using the variants.

Abstract: Systems and methods for performing source separation are provided. Source separation is performed using a composite signal and a signal dictionary. The composite signal is a mixture of sources received by a sensor. The signal dictionary is a database of filtered basis functions that are formed by the application of directional filters. The directional filters approximate how a particular source will be received by the sensor when the source originates from a particular location. Each source can be characterized by a coefficient and a filtered basis function. The coefficients are unknown when the sources are received by the sensor, but can be estimated using the composite signal and the signal dictionary. Various ones of the sources may be selectively reconstructed or separated using the estimated value of the coefficients.