Compositions and methods for targeted depletion, enrichment, and segmentation of nucleic acids using CRISPR/Cas system proteins

• Main Authors: BUSTAMANTE CARLOS D, GOURGUECHON STEPHANE B, CARPENTER MEREDITH L
• Format: Patent
• Language: Chinese; English
• Published: 06.05.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

Provided herein are methods and compositions for depleting a targeted nucleic acid sequence from a sample, enriching a sequence of interest from a sample, and/or segmenting a sequence from a sample. In one aspect, provided herein is a method of enriching a sequence of interest in a sample, comprising: (a) providing a sample comprising a sequence of interest and a sequence targeted to be depleted wherein the sequence of interest comprises less than 30% of the sample; (b) contacting the sample with a plurality of CRISPR/Cas system protein-gRNA complexes wherein the gRNA is complementary to the targeting sequence, and thereby cleaving the targeting sequence. In one embodiment disclosed herein, the method further includes extracting a sequence of interest and a sequence targeted to be subtracted from a sample. 本文提供了用于从样品消减靶向的核酸序列、从样品富集目的序列和/或从样品分割序列的方法和组合物。在一个方面，本文提供了富集样品的目的序列的方法，包括：(a)提供包含目的序列和被靶向以待消减的序列的样品，其中所述目的序列占样品的少于30％；(b)将样品与多个CRISPR/Cas系统蛋白-gRNA复合物接触，其中所述gRNA与靶向序列互补，并由此切割靶向序列。在本文公开的一个实施方案中，该方法进一步包括从样品提取目