# US 10,766,934 B2 · Genetically modified microalgae

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

- Number: US 10,766,934 B2
- Document ID: US10766934B2
- Status: Granted
- Kind: B2
- Application number: 16/086,923
- Filed: 2017-03-10
- Published: 2019-04-11
- Granted: 2020-09-08
- Earliest claimed: 2016-03-20
- Assignees: Reliance Industries Limited
- Applicants: Reliance Industries Limited
- Inventors: Manoj Bhosale; Gautam Das; Tuhin Maity; Pranali Deore; Venkatesh Prasad; Santanu Dasgupta
- Web page: https://patentgrep.com/patent/US10766934B2

## CPC

- C07K 14/405: from algae
- C12N 1/12: Unicellular algae; Culture media therefor (as new plants A01H13/00)
- C12N 15/79: Vectors or expression systems specially adapted for eukaryotic hosts
- C12N 15/82: for plant cells {, e.g. plant artificial chromosomes (PACs)}
- 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)

## Abstract

The present disclosure provides genetically modified Chlorella sorokiniana. DAD1 gene is overexpressed in wild type Chlorella sorokiniana to obtain the genetically modified Chlorella sorokiniana. The genetically modified Chlorella sorokiniana is characterized by increased biomass, increased carrying capacity, reduced clumping, and increased stress tolerance to ultraviolet light and high light intensity, as compared to the wild type Chlorella sorokiniana. The genetically modified Chlorella sorokiniana can be used for the production of industrially important products.

## Claims

### Claim 1

Genetically modified Chlorella sorokiniana having Accession Number CCAP 211/132.

### Claim 2 (depends on 1)

The genetically modified Chlorella sorokiniana as claimed in claim 1, wherein the sequence as set forth in SEQ ID NO. 1 is overexpressed in wild type Chlorella sorokiniana to obtain said genetically modified Chlorella sorokiniana.

### Claim 3 (depends on 1)

The genetically modified Chlorella sorokiniana as claimed in claim 1, wherein said genetically modified Chlorella sorokiniana is characterized by increased biomass, increased carrying capacity, reduced clumping, and increased stress tolerance to ultraviolet light and high light intensity as compared to wild type Chlorella sorokiniana.

### Claim 4 (depends on 2)

The genetically modified Chlorella sorokiniana as claimed in claim 2, wherein said DAD1 gene is obtained from Chlamydomonas reinhardtii NW_001843471 from which the sequence as set forth in SEQ ID NO. 1 is identified and is codon-optimized for Chlorella sorokiniana.

### Claim 5 (depends on 1)

A method of genetically modifying wild type Chlorella sorokiniana to obtain genetically modified Chlorella sorokiniana having Accession Number CCAP 211/132 as claimed in claim 1, said method comprising the following steps:
a. obtaining at least one copy of DAD1 gene from Chlamydomonas reinhardtii NW_001843471 and identifying the sequence as set forth in SEQ ID NO. 1;
b. isolating said SEQ ID NO. 1 from said DAD1 gene and cloning it in a vector, wherein said SEQ ID NO. 1 is codon-optimized for Chlorella sorokiniana; and
c. introducing said vector containing said sequence as set forth in SEQ ID NO. 1 into wild type Chlorella sorokiniana and growing on a medium containing a selective agent under conducive conditions to facilitate the overexpression of said sequence as set forth in SEQ ID NO. 1 to obtain said genetically modified Chlorella sorokiniana having Accession Number CCAP 211/132.

### Claim 6 (depends on 5)

The method as claimed in claim 5, wherein said vector is pChlamy 1.

### Claim 7 (depends on 5)

The method as claimed in claim 5, wherein said selective agent is hygromycin.

### Claim 8 (depends on 5)

The method as claimed in claim 5, wherein the step of cloning is carried out by a technique selected from the group consisting of transposon-mediated insertion, electroporation, magnetophoresis, microinjection, particle inflow gun bombardment, transfection, heat shock transfection, nucleofection transfection, delivery via liposomes and microinjection.