# US 8,367,392 B2 · Genetic transformation of algal and cyanobacteria cells by microporation

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,367,392 B2
- Document ID: US8367392B2
- Status: Granted
- Kind: B2
- Application number: 12/584,571
- Filed: 2009-09-08
- Published: 2010-04-01
- Granted: 2013-02-05
- Earliest claimed: 2008-09-05
- Assignees: TransAlgae Ltd
- Applicants: Daniella Schatz; Doron Eisenstadt; Jonathan Gressel; Shai Ufaz; Ofra Chen; Dikla Journo Eckstein
- Inventors: Daniella Schatz; Doron Eisenstadt; Jonathan Gressel; Shai Ufaz; Ofra Chen; Dikla Journo Eckstein
- Web page: https://patentgrep.com/patent/US8367392B2

## CPC

- C07K 14/62: Insulins
- C12N 1/12: Unicellular algae; Culture media therefor (as new plants A01H13/00)
- C12N 1/20: Bacteria; Culture media therefor
- C12N 9/16: acting on ester bonds (3.1)
- C12N 9/88: Lyases (4.)
- C12N 9/90: Isomerases (5.)
- C12N 9/93: {Ligases (6)}
- C12N 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)}
- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione
- C12Y 401/01039: Ribulose-bisphosphate carboxylase (4.1.1.39)
- C12N 2310/11: Antisense
- C12N 2310/14: interfering nucleic acids [NA]
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES

## Abstract

A method for transforming algal or cyanobacterial cells, comprising mixing a polynucleotide for transforming the cells with the polynucleotide; performing microporation by applying a plurality of electrical pulses to the cells with a microporation apparatus; and incubating said polynucleotide with the cells after said applying said electrical pulses.

## Claims

### Claim 1

A method for transforming algal or cyanobacterial cells, comprising mixing a polynucleotide for transforming the cells with the polynucleotide; performing microporation by applying a plurality of electrical pulses to the cells with a microporation apparatus; and incubating said polynucleotide with the cells after said applying said electrical pulses.

### Claim 2 (depends on 1)

The method of claim 1, wherein said performing microporation by said applying said plurality of electrical pulses comprises applying pulses having a voltage in a range of from 700 to 1700 volts.

### Claim 3 (depends on 2)

The method of claim 2, wherein said applying said pulses comprises applying pulses having a pulse length in a range of from 10 to 40 ms.

### Claim 4 (depends on 3)

The method of claim 3, wherein said applying said plurality of electrical pulses comprises applying said electrical pulses 1-5 times.

### Claim 5 (depends on 4)

The method of claim 4, wherein said mixing comprises suspending the cells in a solution; and mixing said polynucleotide with said solution.

### Claim 6 (depends on 5)

The method of claim 5, wherein said mixing comprises mixing a linear DNA polynucleotide with the cells.

### Claim 7 (depends on 5)

The method of claim 5, wherein said suspending the cells in said solution comprises suspending the cells in a solution that does not remove the cell wall, such that said performing said microporation comprises applying said plurality of electrical pulses to the cells with cell walls.