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Extensive length and homology dependent chimerism in pool-packaged AAV libraries

ABSTRACT: Extensive length and homology dependent chimerism in pool-packaged AAV libraries

PROVIDER: PRJNA1199511 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR31751121_1.fastq.gz	Fastqsanger.gz
	SRR31751122_1.fastq.gz	Fastqsanger.gz
	SRR31751123_1.fastq.gz	Fastqsanger.gz
	SRR31751124_1.fastq.gz	Fastqsanger.gz
	SRR31751125_1.fastq.gz	Fastqsanger.gz

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Extensive length and homology dependent chimerism in pool-packaged AAV libraries

Project description:Adeno-associated viruses (AAVs) have emerged as the foremost gene therapy delivery vehicles due to their versatility, durability and safety profile. Here we demonstrate extensive chimerism among complex AAV libraries that are packaged as a pool. The observed chimerism is length- and homology-dependent but capsid-independent, in some cases affecting >60% of packaged AAV genomes. These results have important implications for the design and deployment of functional AAV libraries in research and clinical settings.

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Extensive length and homology dependent chimerism in pool-packaged AAV libraries.

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Project description:We found that chicken PGCs derived from E7 to E11 gonads were heterogeneity which could cultre in vitro for a long time with germline chimerism and the ratio of chimerism was improved through second blood vessel injection.

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Project description:Two independent ES cell lines (D4 and C2) were assessed for chimerism and germline transmission. Although these lines were able to produce chimerism at early passage, no germ line transmission was seen. After passaging in a proprietary medium and subcloning by detaching loosely connected cells and culturing in the recommended medium, both lines experienced an increase in potential as evidenced by an enhanced ability to go germ line. Analysis of early passage, and late passage detached and attached cells will allow the determination of genes that may be implicated in the increase of potential seen in these cells. Note: additional replicates may be added at a later date. Keywords: embryonic stem cells, chimerism, germline transmission

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Xenophagocytosis blockade enhances interspecies chimerism

Project description:Organ shortage remains a major challenge in transplantation medicine. Interspecies blastocyst complementation offers a promising strategy to generate human organs in livestock. However, efficient xenogeneic donor cell engraftment remains challenging. Here, we identify an innate immune barrier wherein host macrophages selectively eliminate viable xenogeneic donor cells, a process we term xenophagocytosis. Mechanistically, xenogeneic cells display elevated phosphatidylserine, an "eat-me" signal recognized by host macrophages through phagocytic receptor Axl. We demonstrate three orthogonal strategies for xenophagocytosis blockade: genetic ablation of macrophages or Axl receptor in the host embryo, or overexpression of “don’t eat me” signal CD47 or the phosphatidylserine-regulating flippase ATP11C in donor cells. Xenophagocytosis blockade enhances rat and human donor chimerism in mouse embryos and improves interspecies pancreas complementation efficiency. These findings reveal a previously unrecognized innate immune barrier that safeguards species integrity during early embryogenesis and provide mechanistic insights to enhance xenogeneic chimerism for generating human organs in livestock.

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Project description:Interventions: A:JingChang Pre-packaged Product No.1 (4g Konjac Powder);G:JingChang Pre-packaged Product No.2 (2g Konjac Powder);F:JingChang Pre-packaged Product No.3 (1g Konjac Powder);D:Give competing formulations currently available on the market;E:Self prepared diet Primary outcome(s): Clarity of stools after bowel cleansing Study Design: Parallel

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Project description:Two independent ES cell lines (D4 and C2) were assessed for chimerism and germline transmission. Although these lines were able to produce chimerism at early passage, no germ line transmission was seen. After passaging in a proprietary medium and subcloning by detaching loosely connected cells and culturing in the recommended medium, both lines experienced an increase in potential as evidenced by an enhanced ability to go germ line. Analysis of early passage, and late passage detached and attached cells will allow the determination of genes that may be implicated in the increase of potential seen in these cells. Note: additional replicates may be added at a later date. Experiment Overall Design: this experiment include 6 samples and 34 replicates

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