Compositions and methods for targeted depletion, enrichment, and partitioning of nucleic acids using CRISPR/Cas system proteins
Provided herein are methods and compositions for depleting targeted nucleic acid sequences from a sample, enriching for sequences of interest from a sample, and/or partitioning of sequences from a sample. The methods and compositions are applicable to biological, clinical, forensic, and environmental samples.
1. A composition comprising a mixture of DNA targeted sequences for depletion and DNA sequences of interest, wherein the DNA targeted sequences for depletion and the DNA sequences of interest are adapter-ligated, wherein the adapters are ligated to both the 5′ and 3′ ends of both the target sequences for depletion and the sequences of interest, and wherein the DNA targeted sequences for depletion are complexed to a gRNA-CRISPR/Cas system protein complex and wherein the DNA sequences of interest are in the range of about 0.1% to about 50% of the DNA in the composition, wherein the CRISPR/Cas system protein is Cas9, Cpf1, Cas3, Cas8a-c, Cas10, Csel, Csyl, Csn2, Cas4, Csm2, or CmS.
2. The composition of claim 1 , wherein the targeted sequences for depletion are from a host organism and the sequences of interest are from a non-host organism.
3. The composition of claim 1 , wherein the CRISPR/Cas system protein is thermostable.
4. The composition of claim 1 , wherein the composition comprises at least 10 unique CRISPR/Cas system protein-gRNA complexes.
5. The composition of claim 1 , wherein the DNA targeted sequences for depletion and the DNA sequences of interest are from any one of a biological sample, a clinical sample, a forensic sample or an environmental sample.
6. The composition of claim 2 , wherein the non-host organism nucleic acid comprises microbial nucleic acid sequences.
7. The composition of claim 1 , wherein the gRNAs are complementary to mitochondrial DNA.
8. The composition of claim 1 , wherein the gRNAs are complementary to DNA corresponding to ribosomal RNA sequences, sequences encoding globin proteins, sequences encoding a transposon, sequences encoding retroviral sequences, sequences comprising telomere sequences, sequences comprising sub-telomeric repeats, sequences comprising centromeric sequences, sequences comprising intron sequences, sequences comprising Alu repeats, sequences comprising SINE repeats, sequences comprising LINE repeats, sequences comprising dinucleic acid repeats, sequences comprising trinucleic acid repeats, sequences comprising tetranucleic acid repeats, sequences comprising poly-A repeats, sequences comprising poly-T repeats, sequences comprising poly-C repeats, sequences comprising poly-G repeats, sequences comprising AT-rich sequences, or sequences comprising GC-rich sequences.
9. The composition of claim 1 , wherein the DNA targeted sequences for depletion and the DNA sequences of interest are from a sample selected from the group consisting of whole blood, plasma, serum, tears, saliva, mucous, cerebrospinal fluid, teeth, bone, fingernails, feces, urine, tissue, and a biopsy.
10. The composition of claim 2 , wherein the host organism is selected from the group consisting of a human, cow, horse, sheep, pig, monkey, dog, cat, gerbil, bird, mouse, and rat.
11. The composition of claim 2 , wherein the non-host organism is a bacteria, a virus or a eukaryotic parasite or pathogen.
12. The composition of claim 1 , wherein the composition comprises at least 10 2 unique CRISPR/Cas system protein-gRNA complexes.
13. The composition of claim 1 , wherein the adapters are Y-shaped adapters or poly-G adapters.
14. The composition of claim 13 , wherein the adapter-ligated sequences range from 50-1000 bp.