Disclosed are the nucleotide sequences of promoters from Arxula adeninivorans and Yarrowia lipolytica which may be used to drive gene expression in a cell. The promoters were validated, and selected promoters were screened to determine which promoters may be useful for increasing the lipid production efficiency of oleaginous yeasts.

The present invention relates to the field of fungal biotechnology, more particularly to genetic engineering methods for the production of polyunsaturated fatty acids (PUFA) in fungal hosts selected from Rhodosporidium and Rhodotorula genera. The present invention further relates to a modified fungal host cell having reduced native aldehyde dehydrogenase (ALD 1) enzyme activity, and methods for producing omega-3 and omega-6 fatty acids and triacylglycerides, by growing said fungal host cell under suitable conditions.

Disclosed are nucleotide sequences and corresponding amino acid sequences of Arxula adeninivorans genes that can be utilized to manipulate the lipid content and/or composition of a cell. Methods and compositions for utilizing this information are disclosed to increase the lipid content or modify the lipid composition of a cell by either increasing or decreasing the activity of certain genetic targets.

Disclosed are methods and compositions for increasing the triacylglycerol content of a cell by increasing the activity of a type 1 diacylglycerol acyltransferase (i.e., DGA2) and increasing the activity of a type 2 diacylglycerol acyltransferase (i.e., DGA1). In some embodiments, the triacylglycerol content of a cell is also modified by decreasing the activity of a triacylglycerol lipase in the same cell. Also disclosed are methods and compositions for increasing the triacylglycerol content of a cell by increasing the activity of a type 1 diacylglycerol acyltransferase (i.e., DGA2), or by increasing the activity of a type 3 diacylglycerol acyltransferase (i.e., DGA3).

Some aspects provide engineered microbes for oil production. Methods for microbe engineering and for use of engineered microbes are also provided herein. Such engineered microbes exhibit greatly enhanced conversion yields and TAG synthesis and storage properties.

The present invention relates to oleaginous yeast strains overexpressing a hexokinase gene, wherein said strains are capable of accumulating lipids. Methods for obtaining said strains as well as methods for producing lipids are also disclosed.

Recombinant DNA techniques are used to produce oleaginous recombinant cells that produce triglyceride oils having desired fatty acid profiles and regiospecific or stereospecific profiles. Genes manipulated include those encoding stearoyl-ACP desturase, delta 12 fatty acid desaturase, acyl-ACP thioesterase, ketoacyl-ACP synthase, and lysophosphatidic acid acyltransferase. The oil produced can have enhanced oxidative or thermal stability, or can be useful as a frying oil, shortening, roll-in shortening, tempering fat, cocoa butter replacement, as a lubricant, or as a feedstock for various chemical processes. The fatty acid profile can be enriched in midchain profiles or the oil can be enriched in triglycerides of the saturated-unsatturated-saturated type.

Disclosed are methods and compositions for increasing the triacylglycerol content of a cell by up-regulating diacylglycerol acyltransferase and down-regulating triacylglycerol lipase. In some embodiments, a DGA1 protein is expressed and a native TGL3 gene is knocked out, thereby increasing the synthesis of triacylglycerol and decreasing its consumption, respectively.

The present disclosure describes short antisense compounds, including such compounds comprising chemically-modified high-affinity monomers 8-16 monomers in length. Certain such short antisense compound are useful for the reduction of target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, the described short antisense compounds have greater potential for oral dosing.

The invention refers to fungal cells, and especially to oleaginous fungal cells that have been genetically modified to produce enzymes of the pyruvate dehydrogenase bypass route to enhance their lipid production. Especially the cells are modified to overexpress genes encoding pyruvate decarboxylase (PDC), acetaldehyde dehydrogenase (ALD) and/or acetyl-CoA synthetase (ACS), optionally together with a gene encoding diacylglycerol acyltransferase (DAT), or to express genes encoding PDC together with ALD and/or ACS. Methods of producing lipids, biofuels and lubricants using the modified fungi are also disclosed as well as expression cassettes useful therein. A new enzyme having phosholipid:diacylglycerol acyltransferase (PDAT) activity and a polynucleotide encoding it are also disclosed, which are useful in the lipid production. A recombinant Cryptococcus cell and its construction is described.