The specification provides methods for introducing a desired genetic change in a nucleotide sequence using a double-strand break (DSB)-inducing genome editing system, the method comprising: identifying one or more available cut sites in a nucleotide sequence; analyzing the nucleotide sequence and available cut sites with a computational model to identify the optimal cut site for introducing the desired genetic change into the nucleotide sequence; and contacting the nucleotide sequence with a DSB-inducing genome editing system, thereby introducing the desired genetic change in the nucleotide sequence at the cut site.

A method and system for identification of target sites in protein coding regions for combating pathogens has been provided. The method relates to identifying a set of nucleotide repeat sequences that occur within the complete coding region of a specific protein that is involved in the pathogenicity of the infectious bacteria and which occurs in multiple copies on the pathogen genome and utilizing various laboratory acceptable methods to debilitate the identified target sequence on the pathogen genome, as well as use of enzymatic machinery to target and cleave its flanking genes on the genome. The set of nucleotide repeat sequences forming a part of or the complete coding region of a specific protein on the pathogen genome may be flanked by genes on either side that can be targeted as well. The present disclosure further includes administration of a cocktail comprising antimicrobial drugs, biofilm inhibitors and the novel construct.

A method and system for identification of target sites in protein coding regions for combating pathogens has been provided. The method relates to identifying a set of nucleotide repeat sequences that occur within the complete coding region of a specific protein that is involved in the pathogenicity of the infectious bacteria and which occurs in multiple copies on the pathogen genome and utilizing various laboratory acceptable methods to debilitate the identified target sequence on the pathogen genome, as well as use of enzymatic machinery to target and cleave its flanking genes on the genome. The set of nucleotide repeat sequences forming a part of or the complete coding region of a specific protein on the pathogen genome may be flanked by genes on either side that can be targeted as well. The present disclosure further includes administration of a cocktail comprising antimicrobial drugs, biofilm inhibitors and the novel construct.

Provided herein are compositions and methods for preserving proximity data in nucleic acid samples, by embedding indexing information in the samples prior to fragmentation. Further provided herein are transposon libraries for generating such indexed nucleic acid samples.

This disclosure relates to methods of generating antibody libraries, antibody libraries produced using such methods, and variant antibodies. Presently, methods of improving antibody binding (affinity maturation assays) require the screening of vast libraries of antibody variants (often >10.sup.10) to identify a small fraction of variants with improved characteristics. The present invention involves taking the nucleotide sequence of the framework and complementarity determining region of a target antibody and identifying motifs which would be recognised by deamination somatic hypermutation enzymes. A small library of variants is then created which incorporate one or more of these mutations. It was found that a relatively high proportion of the variants have an increased affinity. The technique of the present invention was demonstrated on the trastuzumab and Cathepsin S antibodies, and the variants produced are also claimed.

Aspects of the invention relate to methods, compositions and algorithms for designing and producing a target nucleic acid. The method can include: (1) providing a plurality of blunt-end double-stranded nucleic acid fragments having a restriction enzyme recognition sequence at both ends thereof; (2) producing via enzymatic digestion a plurality of cohesive-end double-stranded nucleic acid fragments each having two different and non-complementary overhangs; (3) ligating the plurality of cohesive-end double-stranded nucleic acid fragments with a ligase; and (4) forming a linear arrangement of the plurality of cohesive-end double-stranded nucleic acid fragments, wherein the unique arrangement comprises the target nucleic acid. In certain embodiments, the plurality of blunt-end double-stranded nucleic acid fragments can be provided by: releasing a plurality of oligonucleotides synthesized on a solid support; and synthesizing complementary strands of the plurality of oligonucleotides using a polymerase based reaction.

A method for predictive engineering of a sample derived from a genetically optimized non-human donor suitable for xenotransplantation into a human having improved quality or performance is provided. The method includes constructing a training data set from a series of libraries, wherein at least one library in the series of libraries comprises genomic, proteomic, and research data specific to non-humans. The method includes developing a predictive machine learning model based on the constructed training data set. The method includes utilizing the predictive machine learning model to obtain a predicted quality or performance of a plurality of sequences for a candidate sample from the non-human donor specific to a human patient or patient population. The method includes selecting a subset of sequences for evaluation from the plurality of sequences based on the predicted quality or performance. The method includes designing candidate samples derived from the non-human donor using the selected subset of sequences. The method includes measuring a respective in silico performance of each designed candidate sample. The method includes selecting a designed candidate sample for manufacture based on the respective in silico performance of each designed candidate sample.

Methods and systems for designing, testing, and validating genome designs based on rules or constraints or conditions or parameters or features and scoring are described herein. A computer-implemented method includes receiving data for a known genome and a list of alleles, identifying and removing occurrences of each allele in the known genome, determining a plurality of allele choices with which to replace occurrences in the known genome, generating a plurality of alternative gene sequences for a genome design based on the known genome, wherein each alternative gene sequence comprises a different allele choice, applying a plurality of rules or constraints or conditions or parameters or features to each alternative gene sequence by assigning a score for each rule or constraint or condition or parameter or feature in each alternative gene sequence, resulting in scores for the applied plurality of rules or constraints or conditions or parameters or features, scoring each alternative gene sequence based on a weighted combination of the scores for the plurality of rules or constraints or conditions or parameters or features, and selecting at least one alternative gene sequence as the genome design based on the scoring.

Ordered assembly of large numbers of fragments into a single large DNA have been improved in both frequency and fidelity of the assembled product. This has been achieved by novel compositions and methods that are utilized in a computer system that integrates comprehensive ligation data from multiple sources to provide optimized synthetic overhangs or overhangs from restriction endonuclease cleavage on DNA fragments for assembly by ligation. Intragenic cut sites are avoided by the use of a novel restriction endonuclease which recognizes 7 nucleotides (bases) and cuts DNA to create 4-base overhangs with the help of a synthetic activator oligonucleotide. Variations in ligation preferences by different ligases provide extra precision in assembly reactions. The use of the improved methods are exemplified by the successful assembly from 52 fragments of a viral genome and also a 52 fragment ordered assembly of a bacteria operon.

Embodiments disclosed herein are directed to engineered CRISPR-Cas effector proteins that comprise at least one modification compared to an unmodified CRISPR-Cas effector protein that enhances binding of the of the CRISPR complex to the binding site and/or alters editing preference as compared to wild type. In certain example embodiments, the CRISPR-Cas effector protein is a Type V effector protein. In certain other example embodiments, the Type V effector protein is Cpf1. Embodiments disclosed herein are directed to viral vectors for delivery of CRISPR-Cas effector proteins, including Cpf1. In certain example embodiments, the vectors are designed so as to allow packaging of the CRISPR-Cas effector protein within a single vector. There is also an increased interest in the design of compact promoters for packing and thus expressing larger transgenes for targeted delivery and tissue-specificity. Thus, in another aspect certain embodiments disclosed herein are directed to delivery vectors, constructs, and methods of delivering larger genes for systemic delivery.