De novo synthesized nucleic acid libraries
AssigneeTwist Bioscience Corporation
InventorsAnthony Cox, Siyuan Chen
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Abstract
Disclosed herein are methods for the generation of nucleic acid libraries encoding for gRNA sequences. The gRNAs encoded by methods described herein may be single or double gRNA sequences. Methods described provide for the generation of gRNA libraries, as a DNA precursor or as a RNA transcription product, with improved accuracy and uniformity.
Claims26
A nucleic acid library, wherein the nucleic acid library comprises at least 500 non-identical DNA molecules, wherein each non-identical DNA molecule encodes for a different gRNA sequence, wherein each gRNA sequence comprises a targeting domain complementary to a mammalian gene, and wherein at least about 80% of the at least 500 non-identical DNA molecules are each present in the nucleic acid library in an amount within 2× of a mean frequency for each of the non-identical DNA molecules in the library.
A nucleic acid library, wherein the nucleic acid library comprises at least 2000 non-identical nucleic acids, wherein each non-identical nucleic acid encodes for a different gRNA sequence, wherein each gRNA sequence comprises a targeting domain complementary to a eukaryotic gene, and wherein at least about 80% of the at least 2000 non-identical nucleic acids are present in the nucleic acid library in an amount within 2× of a mean frequency for each of the non-identical nucleic acids in the library.
An amplicon library, wherein the amplicon library comprises a plurality of non-identical DNA molecules, wherein each non-identical DNA molecule is present in a population of amplification products, wherein each non-identical DNA molecule encodes for a different gRNA sequence, wherein each gRNA sequence comprises a targeting domain complementary to a eukaryotic gene, and wherein at least about 80% of the plurality of non-identical DNA molecules are each present in the amplicon library in an amount within 2× of a mean frequency for each of the non-identical DNA molecules in the library.
Dependencies are inferred from the claim text.