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1.
Nat Commun ; 13(1): 1028, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35232966

RESUMO

Prime editing is a versatile genome-editing technique that shows great promise for the generation and repair of patient mutations. However, some genomic sites are difficult to edit and optimal design of prime-editing tools remains elusive. Here we present a fluorescent prime editing and enrichment reporter (fluoPEER), which can be tailored to any genomic target site. This system rapidly and faithfully ranks the efficiency of prime edit guide RNAs (pegRNAs) combined with any prime editor variant. We apply fluoPEER to instruct correction of pathogenic variants in patient cells and find that plasmid editing enriches for genomic editing up to 3-fold compared to conventional enrichment strategies. DNA repair and cell cycle-related genes are enriched in the transcriptome of edited cells. Stalling cells in the G1/S boundary increases prime editing efficiency up to 30%. Together, our results show that fluoPEER can be employed for rapid and efficient correction of patient cells, selection of gene-edited cells, and elucidation of cellular mechanisms needed for successful prime editing.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Genoma , Humanos , Mutação , RNA Guia de Cinetoplastídeos/genética
2.
J Dairy Sci ; 99(3): 1747-1754, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26805991

RESUMO

Sixty percent of milk consumed in China has a long shelf life (UHT), presumably because milk with a short shelf life (pasteurized) is comparatively expensive. This in contrast to Australia, where 10% of consumed milk is UHT and the price between UHT and pasteurized milk is equivalent. Whether UHT is actually more liked than pasteurized milk by Chinese consumers is unknown. However, the potential positive halo around the expensive pasteurized milk might result in Chinese consumers liking milk more when it is labeled as "short shelf-life milk." To test these hypotheses, Chinese (n=48, 20 males, 28 females, 23 ± 7.2 yr) and Australian (n=93, 11 males, 82 females, 24 ± 5.6 yr) consumers tasted and rated (9-point hedonic scale), in a randomized order, 3 × 30-mL samples of UHT milk (labeled as "long shelf-life milk," "short shelf-life milk," or "milk") and 3 × 30-mL samples of pasteurized milk (also labeled as "long shelf-life milk," "short shelf-life milk," or "milk"). Australian participants' liking of milk was not influenced by labeling. Regardless of what the label stated, they always preferred the taste of pasteurized milk over the taste of UHT milk. This was different for Chinese participants, who preferred the taste of UHT milk over the taste of pasteurized milk, but in general had a higher liking for any milk that was labeled "short shelf-life milk." Both Australian and Chinese were more positive about pasteurized than UHT milk. In conclusion, Chinese, but not Australian, consumers' liking of milk was guided by the positive expectations of pasteurized milk and the negative expectations of UHT milk. Further research is needed to investigate if the present findings can be extrapolated to a larger and more varied group of Chinese and Australian consumers.


Assuntos
Bovinos/fisiologia , Rotulagem de Alimentos , Preferências Alimentares , Leite , Animais , Austrália , China , Comportamento do Consumidor , Feminino , Manipulação de Alimentos , Humanos , Masculino , Pasteurização , Paladar
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