Abstract: Provided herein are safe harbor loci that can be utilized as sites for genetic modification. Safe harbor loci of the disclosure are shown to support sustained transgene expression with minimal silencing, and minimal impact on local or global gene expression. Safe harbor loci disclosed herein can be used in various genetic and cell engineering applications.

Abstract: Described herein are compositions and methods for modifying and targeting genes. Also described herein are compositions and methods for modifying and targeting genes in a cell or a non-human mammal.

Abstract: The present disclosure describes systems and methods for immunotherapies Immune cells can be engineered to exhibit enhanced half-life as compared to control cell (e.g., a non-engineered immune cell). Immune cells can be engineered to exhibit enhanced proliferation as compared to a control cell. Immune cells can be engineered to effectively and specifically target diseased cells (e.g., cancer cells) that a control cell otherwise is insufficient or unable to target. The engineered Immune cells disclosed herein can be engineered ex vivo, in vitro, and in some cases, in vivo. The engineered Immune cells that are prepared ex vivo or in vitro can be administered to a subject in need thereof to treat a disease (e.g., myeloma or solid tumors). The engineered Immune cells can be autologous to the subject. Alternatively, the engineered immune cells can be allogeneic to the subject.

Abstract: Described here are methods, compositions, and systems for generating transgenic islet cells suitable for xenotransplantation.

Abstract: Provided are cells, tissues, organs, and/or animals having one or more modified genes for enhanced xenograft survival and/or tolerance. And methods of producing and using the cells, tissues, organs, and/or animals.

Abstract: The present disclosure generally relates to nucleic acid amplification systems and methods suitable for construction of nucleic acid samples, including construction of normalized nucleic acid libraries. In some embodiments, the method includes providing one or more input nucleic acid samples, contacting each of the input nucleic acid samples (e.g., input library) with a reaction mixture including first amplification or normalization primers and second amplification or normalization primers, wherein the first amplification or normalization primers are immobilized on a solid support and the second amplification or normalization primers are in solution phase, and amplifying the input nucleic acid samples under conditions such that substantially all of the first amplification or normalization primers are incorporated into amplification products. Further provided are systems and droplet actuator devices that are configured to carry out the methods disclosed herein.

Abstract: The disclosure provides methods of DNA amplification mediated by DNA ligase. Specifically disclosed is a method of amplifying a target region at DNA level, which comprises repeating cycles of amplification comprising the following steps: denaturing DNA template to obtain single target DNA strands; hybridizing a primer pair comprising an upstream primer and a downstream primer to the target DNA strand, wherein the upstream primer is hybridized to a first nucleic acid sequence of the target region, and the downstream primer is hybridized to a second nucleic acid sequence of the target region; the first nucleic acid sequence is downstream to the second nucleic acid sequence on the target DNA strand, with the downstream primer containing a phosphorylated 5? end; ligating the upstream primer or extension product thereof to the downstream primer or extension product thereof, to obtain a semi-amplification product. This disclosure also discloses a kit used to amplify a target region at DNA level.

Abstract: The present disclosure generally relates to nucleic acid amplification systems and methods suitable for construction of nucleic acid samples, including construction of normalized nucleic acid libraries. In some embodiments, the method includes providing one or more input nucleic acid samples, contacting each of the input nucleic acid samples (e.g., input library) with a reaction mixture including first amplification or normalization primers and second amplification or normalization primers, wherein the first amplification or normalization primers are immobilized on a solid support and the second amplification or normalization primers are in solution phase, and amplifying the input nucleic acid samples under conditions such that substantially all of the first amplification or normalization primers are incorporated into amplification products. Further provided are systems and droplet actuator devices that are configured to carry out the methods disclosed herein.

Abstract: The present application discloses a method of detecting the methylation of specific genome regions in a DNA fragment from a sample containing DNA. The method includes treating the DNA fragment containing specific genome regions with sodium bisulfite and obtaining single-strand DNA fragment; attaching an adapter to one or both ends of the single-strand DNA fragment; optionally cyclizing the adapter-attached single-strand DNA fragment; preparing the single-strand DNA fragment with attached adapter into a DNA sequencing library containing the specific genome regions; sequencing the DNA sequencing library to identify the sequence of the single-strand DNA fragment. The present application also discloses a kit for detecting the methylation of specific genome regions in a DNA fragment from a sample containing DNA, and the use of single strand ligating agent in preparing a kit for detecting the methylation of specific genome regions in a DNA fragment.