diff --git a/packages/env/lib/config/test/config.js b/packages/env/lib/config/test/config.js
index 5a79cb6d4f724a..1d5060f11ed799 100644
--- a/packages/env/lib/config/test/config.js
+++ b/packages/env/lib/config/test/config.js
@@ -4,6 +4,7 @@
  */
 const { readFile, stat } = require( 'fs' ).promises;
 const os = require( 'os' );
+const { join, resolve } = require( 'path' );
 
 /**
  * Internal dependencies
@@ -101,13 +102,17 @@ describe( 'readConfig', () => {
 			process.env.WP_ENV_HOME = 'here/is/a/path';
 			const configWith = await readConfig( '.wp-env.json' );
 			expect(
-				configWith.workDirectoryPath.includes( 'here/is/a/path' )
+				configWith.workDirectoryPath.includes(
+					join( 'here', 'is', 'a', 'path' )
+				)
 			).toBe( true );
 
 			process.env.WP_ENV_HOME = undefined;
 			const configWithout = await readConfig( '.wp-env.json' );
 			expect(
-				configWithout.workDirectoryPath.includes( 'here/is/a/path' )
+				configWithout.workDirectoryPath.includes(
+					join( 'here', 'is', 'a', 'path' )
+				)
 			).toBe( false );
 
 			process.env.WP_ENV_HOME = oldEnvHome;
@@ -126,13 +131,17 @@ describe( 'readConfig', () => {
 			process.env.WP_ENV_HOME = 'here/is/a/path';
 			const configWith = await readConfig( '.wp-env.json' );
 			expect(
-				configWith.workDirectoryPath.includes( 'here/is/a/path' )
+				configWith.workDirectoryPath.includes(
+					join( 'here', 'is', 'a', 'path' )
+				)
 			).toBe( true );
 
 			process.env.WP_ENV_HOME = undefined;
 			const configWithout = await readConfig( '.wp-env.json' );
 			expect(
-				configWithout.workDirectoryPath.includes( 'here/is/a/path' )
+				configWithout.workDirectoryPath.includes(
+					join( 'here', 'is', 'a', 'path' )
+				)
 			).toBe( false );
 
 			process.env.WP_ENV_HOME = oldEnvHome;
@@ -242,26 +251,31 @@ describe( 'readConfig', () => {
 			readFile.mockImplementation( () =>
 				Promise.resolve(
 					JSON.stringify( {
-						plugins: [ './relative', '../parent', '~/home' ],
+						plugins: [
+							'./relative',
+							'../parent',
+							`${ os.homedir() }/home`,
+						],
 					} )
 				)
 			);
 			const config = await readConfig( '.wp-env.json' );
+
 			expect( config.env.development ).toMatchObject( {
 				pluginSources: [
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*relative$/ ),
+						path: expect.stringMatching( /^(\/||\\).*relative$/ ),
 						basename: 'relative',
 					},
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*parent$/ ),
+						path: expect.stringMatching( /^(\/||\\).*parent$/ ),
 						basename: 'parent',
 					},
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*home$/ ),
+						path: expect.stringMatching( /^(\/||\\).*home$/ ),
 						basename: 'home',
 					},
 				],
@@ -270,17 +284,17 @@ describe( 'readConfig', () => {
 				pluginSources: [
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*relative$/ ),
+						path: expect.stringMatching( /^(\/||\\).*relative$/ ),
 						basename: 'relative',
 					},
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*parent$/ ),
+						path: expect.stringMatching( /^(\/||\\).*parent$/ ),
 						basename: 'parent',
 					},
 					{
 						type: 'local',
-						path: expect.stringMatching( /^\/.*home$/ ),
+						path: expect.stringMatching( /^(\/||\\).*home$/ ),
 						basename: 'home',
 					},
 				],
@@ -310,28 +324,28 @@ describe( 'readConfig', () => {
 			expect( config.env.development.pluginSources ).toEqual( [
 				{
 					type: 'local',
-					path: expect.stringMatching( /^\/.*test1a$/ ),
+					path: expect.stringMatching( /^(\/||\\).*test1a$/ ),
 					basename: 'test1a',
 				},
 			] );
 			expect( config.env.development.themeSources ).toEqual( [
 				{
 					type: 'local',
-					path: expect.stringMatching( /^\/.*test2a$/ ),
+					path: expect.stringMatching( /^(\/||\\).*test2a$/ ),
 					basename: 'test2a',
 				},
 			] );
 			expect( config.env.tests.pluginSources ).toEqual( [
 				{
 					type: 'local',
-					path: expect.stringMatching( /^\/.*test1b$/ ),
+					path: expect.stringMatching( /^(\/||\\).*test1b$/ ),
 					basename: 'test1b',
 				},
 			] );
 			expect( config.env.tests.themeSources ).toEqual( [
 				{
 					type: 'local',
-					path: expect.stringMatching( /^\/.*test2b$/ ),
+					path: expect.stringMatching( /^(\/||\\).*test2b$/ ),
 					basename: 'test2b',
 				},
 			] );
@@ -345,15 +359,19 @@ describe( 'readConfig', () => {
 			expect( config.env.development ).toMatchObject( {
 				coreSource: {
 					type: 'local',
-					path: expect.stringMatching( /^\/.*relative$/ ),
-					testsPath: expect.stringMatching( /^\/.*tests-relative$/ ),
+					path: expect.stringMatching( /^(\/||\\).*relative$/ ),
+					testsPath: expect.stringMatching(
+						/^(\/||\\).*tests-relative$/
+					),
 				},
 			} );
 			expect( config.env.tests ).toMatchObject( {
 				coreSource: {
 					type: 'local',
-					path: expect.stringMatching( /^\/.*relative$/ ),
-					testsPath: expect.stringMatching( /^\/.*tests-relative$/ ),
+					path: expect.stringMatching( /^(\/||\\).*relative$/ ),
+					testsPath: expect.stringMatching(
+						/^(\/||\\).*tests-relative$/
+					),
 				},
 			} );
 		} );
@@ -378,21 +396,21 @@ describe( 'readConfig', () => {
 						type: 'git',
 						url: 'https://github.com/WordPress/gutenberg.git',
 						ref: 'master',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
 						type: 'git',
 						url: 'https://github.com/WordPress/gutenberg.git',
 						ref: 'trunk',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
 						type: 'git',
 						url: 'https://github.com/WordPress/gutenberg.git',
 						ref: '5.0',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
@@ -401,7 +419,7 @@ describe( 'readConfig', () => {
 							'https://github.com/WordPress/theme-experiments.git',
 						ref: 'tt1-blocks@0.4.3',
 						path: expect.stringMatching(
-							/^\/.*theme-experiments\/tt1-blocks$/
+							/^(\/||\\).*theme-experiments(\/||\\)tt1-blocks$/
 						),
 						basename: 'tt1-blocks',
 					},
@@ -431,28 +449,32 @@ describe( 'readConfig', () => {
 						type: 'zip',
 						url:
 							'https://downloads.wordpress.org/plugin/gutenberg.zip',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://downloads.wordpress.org/plugin/gutenberg.8.1.0.zip',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://downloads.wordpress.org/theme/twentytwenty.zip',
-						path: expect.stringMatching( /^\/.*twentytwenty$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*twentytwenty$/
+						),
 						basename: 'twentytwenty',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://downloads.wordpress.org/theme/twentytwenty.1.3.zip',
-						path: expect.stringMatching( /^\/.*twentytwenty$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*twentytwenty$/
+						),
 						basename: 'twentytwenty',
 					},
 				],
@@ -482,34 +504,42 @@ describe( 'readConfig', () => {
 						type: 'zip',
 						url:
 							'https://www.example.com/test/path/to/gutenberg.zip',
-						path: expect.stringMatching( /^\/.*gutenberg$/ ),
+						path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 						basename: 'gutenberg',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://www.example.com/test/path/to/gutenberg.8.1.0.zip',
-						path: expect.stringMatching( /^\/.*gutenberg.8.1.0$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*gutenberg.8.1.0$/
+						),
 						basename: 'gutenberg.8.1.0',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://www.example.com/test/path/to/twentytwenty.zip',
-						path: expect.stringMatching( /^\/.*twentytwenty$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*twentytwenty$/
+						),
 						basename: 'twentytwenty',
 					},
 					{
 						type: 'zip',
 						url:
 							'https://www.example.com/test/path/to/twentytwenty.1.3.zip',
-						path: expect.stringMatching( /^\/.*twentytwenty.1.3$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*twentytwenty.1.3$/
+						),
 						basename: 'twentytwenty.1.3',
 					},
 					{
 						type: 'zip',
 						url: 'https://example.com/twentytwenty.1.3.zip',
-						path: expect.stringMatching( /^\/.*twentytwenty.1.3$/ ),
+						path: expect.stringMatching(
+							/^(\/||\\).*twentytwenty.1.3$/
+						),
 						basename: 'twentytwenty.1.3',
 					},
 				],
@@ -550,12 +580,12 @@ describe( 'readConfig', () => {
 			const matchObj = {
 				test: {
 					type: 'local',
-					path: expect.stringMatching( /^\/.*relative$/ ),
+					path: expect.stringMatching( /^(\/||\\).*relative$/ ),
 					basename: 'relative',
 				},
 				test2: {
 					type: 'git',
-					path: expect.stringMatching( /^\/.*gutenberg$/ ),
+					path: expect.stringMatching( /^(\/||\\).*gutenberg$/ ),
 					basename: 'gutenberg',
 				},
 			};
@@ -653,24 +683,25 @@ describe( 'readConfig', () => {
 			expect( config.env.development.mappings ).toEqual( {
 				test1: {
 					basename: 'test1',
-					path: '/test1',
+					// resolve is required to remove drive letters on Windows.
+					path: resolve( '/test1' ),
 					type: 'local',
 				},
 				test3: {
 					basename: 'test3',
-					path: '/test3',
+					path: resolve( '/test3' ),
 					type: 'local',
 				},
 			} );
 			expect( config.env.tests.mappings ).toEqual( {
 				test1: {
 					basename: 'test1',
-					path: '/test1',
+					path: resolve( '/test1' ),
 					type: 'local',
 				},
 				test2: {
 					basename: 'test2',
-					path: '/test2',
+					path: resolve( '/test2' ),
 					type: 'local',
 				},
 			} );
@@ -702,14 +733,14 @@ describe( 'readConfig', () => {
 			expect( config.env.development.mappings ).toEqual( {
 				test: {
 					basename: 'test3',
-					path: '/test3',
+					path: resolve( '/test3' ),
 					type: 'local',
 				},
 			} );
 			expect( config.env.tests.mappings ).toEqual( {
 				test: {
 					basename: 'test2',
-					path: '/test2',
+					path: resolve( '/test2' ),
 					type: 'local',
 				},
 			} );
diff --git a/packages/scripts/CHANGELOG.md b/packages/scripts/CHANGELOG.md
index 45e0edb0064695..41440738212cb6 100644
--- a/packages/scripts/CHANGELOG.md
+++ b/packages/scripts/CHANGELOG.md
@@ -6,6 +6,10 @@
 
 -   Enable by default code formatting for JSON files in the `format` command ([#40994](https://github.com/WordPress/gutenberg/pull/40994)). You can opt-out of this behavior by providing a custom file matcher, example: `wp-scripts format src/**/*.js`.
 
+### Bug Fixes
+
+-   Fix: env unit test fails on Windows ([#41070](https://github.com/WordPress/gutenberg/pull/41070))
+
 ## 23.0.0 (2022-05-04)
 
 ### Breaking Changes