two items with same first author and year

Exported Items222.json

[
	{
		"id": "http://zotero.org/users/3948081/items/WYEZIQGL",
		"type": "article-journal",
		"abstract": "Transcriptional perturbation using inactivated CRISPR-nucleases (dCas) is a common method in eukaryotic organisms. While rare examples of dCas9-based tools for prokaryotes have been described, multiplexing approaches are limited due to the used effector nuclease. For the first time, a dCas12a derived tool for the targeted activation and repression of genes was developed. Therefore, a previously described SoxS activator domain was linked to dCas12a to enable the programmable activation of gene expression. A proof of principle of transcriptional regulation was demonstrated on the basis of fluorescence reporter assays using the alternative host organism Paenibacillus polymyxa as well as Escherichia coli. Single target and multiplex CRISPR interference targeting the exopolysaccharide biosynthesis of P. polymyxa was shown to emulate polymer compositions of gene knockouts. The simultaneous expression of 11 gRNAs targeting multiple lactate dehydrogenases and a butanediol dehydrogenase resulted in decreased lactate formation, as well as an increased butanediol production in microaerobic fermentation processes. Even though Cas12a is more restricted in terms of its genomic target sequences compared to Cas9, its ability to efficiently process its own guide RNAs in vivo makes it a promising tool to orchestrate sophisticated genetic reprogramming of bacterial cells or to screen for engineering targets in the genome. The developed tool will accelerate metabolic engineering efforts in the alternative host organism P. polymyxa and might be also applied for other bacterial cell factories.",
		"container-title": "ACS Synth Biol",
		"DOI": "10.1021/acssynbio.0c00424",
		"ISSN": "2161-5063 (Electronic) 2161-5063 (Linking)",
		"issue": "12",
		"page": "3353-3363",
		"title": "Novel Prokaryotic CRISPR-Cas12a-Based Tool for Programmable Transcriptional Activation and Repression",
		"volume": "9",
		"author": [
			{
				"family": "Schilling",
				"given": "C."
			},
			{
				"family": "Koffas",
				"given": "M. A. G."
			},
			{
				"family": "Sieber",
				"given": "V."
			},
			{
				"family": "Schmid",
				"given": "J."
			}
		],
		"issued": {
			"date-parts": [
				[
					"2020",
					1
				]
			]
		}
	},
	{
		"id": "http://zotero.org/users/3948081/items/QE8Q24Y9",
		"type": "article-journal",
		"abstract": "Functional carbohydrate polymers are of immense industrial interest for high value applications. Distinct biosynthetic pathways allow for metabolic engineering approaches for production in microbial cell factories. The most common strategies in recent years included the attenuation of central carbon metabolism, improved substrate utilization or enhanced intracellular sugar nucleotide precursor levels. Recombinant expression in more suitable surrogate host organisms has demonstrated remarkable results for the heterologous production of glycosaminoglycans. However, industrial application of pharmacological active functional polysaccharides is often limited by costly post-polymerization modifications and downstream processing. With increasing knowledge of bottleneck enzymes and fluxes, it will be possible to enable a sustainable microbial production of high value polysaccharides and tailor artificial polymers towards specific applications.",
		"container-title": "Curr Opin Biotechnol",
		"DOI": "10.1016/j.copbio.2020.06.010",
		"ISSN": "1879-0429 (Electronic) 0958-1669 (Linking)",
		"page": "44-51",
		"title": "Metabolic engineering for production of functional polysaccharides",
		"volume": "66",
		"author": [
			{
				"family": "Schilling",
				"given": "C."
			},
			{
				"family": "Badri",
				"given": "A."
			},
			{
				"family": "Sieber",
				"given": "V."
			},
			{
				"family": "Koffas",
				"given": "M."
			},
			{
				"family": "Schmid",
				"given": "J."
			}
		],
		"issued": {
			"date-parts": [
				[
					"2020",
					7
				]
			]
		}
	}
]